DOI: 10.5061/dryad.79cnp5j7c
title: Histone acetyltransferase p300/CBP regulates reproductive diapause via the juvenile hormone pathway in the cabbage beetle, Colaphellus bowringi

description: Juvenile hormone (JH) serves as a pivotal endocrine regulator of
 reproductive diapause in insects, though the molecular mechanisms
 governing JH signaling remain poorly characterized. This investigation
 assesses the roles of the histone acetyltransferase p300/CBP in mediating
 reproductive diapause through JH signaling in the cabbage beetle
 Colaphellus bowringi. RNA interference-mediated suppression of p300/CBP in
 reproductive female adults induced diapause-like phenotypes, including
 arrested ovarian maturation, suppressed vitellogenesis, enhanced lipid
 storage, and upregulated expression of diapause-associated genes. Along
 with downregulation of key JH biosynthetic enzymes and signaling
 components. While exogenous JH application decreased lipid accumulation,
 it failed to rescue ovarian developmental arrest. Parallel experiments in
 diapausing females with p300/CBP knockdown yielded similar results - JH
 analog treatment reduced lipid reserves without stimulating ovarian
 development. These findings collectively demonstrate that p300/CBP
 regulates photoperiodic reproductive diapause through dual regulation of
 JH biosynthesis and signal transduction pathways in C. bowringi. Our
 results establish p300/CBP as a critical mediator of JH biosynthesis and
 signaling during photoperiod-induced reproductive diapause. This work
 advances our understanding of epigenetic regulation in insect diapause and
 provides a molecular framework for future studies on environmental
 adaptation strategies in insects.
description: # Histone acetyltransferase p300/CBP regulates reproductive diapause via
 the juvenile hormone pathway in the cabbage beetle, Colaphellus bowringi
 ## General information This dataset does not include large-scale datasets
 such as transcriptomes; instead, it is organized according to the figures
 and tables in the article. It primarily comprises RT-qPCR data and various
 biochemical assay results. The calculated data from RT-qPCR, TG, TL, WB,
 ovarian size measurements, lipid droplet size measurements, and HPLC/MSMS
 are provided in CSV format, while other incompatible data are presented in
 PPT files. Additionally, anatomical images of the cabbage beetle ovaries
 are included. **Corresponding author information** Name: Xiao-Ping Wang
 ORCID:
 [https://orcid.org/0000-0002-1718-1026](https://orcid.org/0000-0002-1718-1026) Affiliation: Hubei Key Laboratory of Insect Resources Utilization and Sustainable Pest Management, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China. email: [xpwang@mail.hzau.edu.cn](mailto:xpwang@mail.hzau.edu.cn) **Alternative contact information** Name: Hao-Min An ORCID: [https://orcid.org/0009-0002-3615-598X](https://orcid.org/0009-0002-3615-598X) Affiliation: Hubei Key Laboratory of Insect Resources Utilization and Sustainable Pest Management, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China. Institute for Advanced Study, Shenzhen University, Shenzhen 518060, China. email: [haominan@szu.edu.cn](mailto:haominan@szu.edu.cn), [ahm1994@126.com](mailto:ahm1994@126.com) **Funding information** This work was supported by National Natural Science Foundation of China (NSFC) grants (32172391). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. # Dryad dataset Dataset DOI: [10.5061/dryad.79cnp5j7c](10.5061/dryad.79cnp5j7c) ## Description of the data and file structure We used dsRNA to knock down the key lysine acetyltransferase p300/CBP gene in the cabbage beetle (*Colaphellus bowringi*), a Coleopteran vegetable pest that undergoes reproductive diapause induced by a long photoperiod. By measuring ovarian phenotypes, lipid accumulation levels, juvenile hormone (JH) synthesis, and JH response levels, we analyzed the mechanism by which p300/CBP regulates the occurrence of reproductive diapause in the cabbage beetle through the JH pathway. Additionally, we combined methoprene induction experiments to analyze the specificity of p300/CBP in regulating the ovarian response to JH signaling in the cabbage beetle. SD means short-day, reproductive females. LD means long-day, diapause females. MG means mid-gut, FB means fat bodies, OVA means ovaries. Fig 1c is a schematic diagram of the RNAi target region and contains no actual data; therefore, it is not included in this dataset. ### Files and variables #### File: IMB_all_data.zip #### File names When applicable, file names follow the pattern `eut.(fig1).R(fig2).(fig3).(fig4).(fig5).(file_extension)`, where: * `(fig1)` * `fig1a_Temporal_expression_pattern.csv` * `group` for different group include heads of LD-0dA-head,SD-0dA-head,LD-1dA-head,SD-1dA-head,LD-2dA-head,SD-2dA-head,LD-3dA-head,SD-3dA-head,LD-4dA-head SD-4dA-head * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of CBP` for ct value of CBP * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation * `fig1b_Tissue_expression_pattern.CSV` * `group` for different group include SD-2dA-MG,SD-2dA-head,SD-2dA-FB,SD-2dA-OVA * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of CBP` for ct value of CBP * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation * `fig1d_SD+P-dsCBP_Mortality.csv` * `group` for different group include SD+P-dsGFP1,SD+P-dsGFP2,SD+P-dsGFP3,SD+P-dsCBP1,SD+P-dsCBP2,SD+P-dsCBP3 * `death number` for count death number * `Mortality %` for mortality calculation * `fig1e_SD_0dA_dsCBP_Survival_rate.CSV` * `n=30` means each group include 30 individuals * `days after RNAi` means days after dsRNA injection * `SD+dsGFP` means dsGFP injection control * `SD+dsCBP` means dsCBP injection * `fig1f_SD+dsCBP_RNAi_efficiency.CSV` * `group` for different group include 4-day-old female adults whole bodies of SD+dsGFP,SD+dsCBP * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of CBP` for ct value of CBP * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation * `fig1g_ovary_class.csv` * `group` for different group include SD+0dA+dsGFP,SD+0dA+dsCBP-F1,SD+0dA+dsCBP-F2 * `class` for different developmental class of ovaries include class 0,Class I,Class II,Class III,Class IV * `Standard` The criteria for classifying ovarian stages. Undifferentiated ovarian tubules lacking vitellogenin deposition=class 0,Ovarian tubule enlargement without vitellogenin deposition=Class I,Expanded ovarian tubules containing vitellogenin deposits but lacking defined eggs=Class II,Developed ovarian tubules with vitellogenin deposits and distinct immature eggs (spherical, size-variable precursors)=Class III,Fully matured ovarian tubules packed with uniform oval-shaped eggs=Class IV * `numbers` numbers of ovaries of class 0,Class I,Class II,Class III and Class IV * `%` Percentage of class 0,Class I,Class II,Class III oand Class IV * `fig1h_SD+dsCBP_ovary_phenotype.zip` include ovary phenotype pics and ovary size sft format files of SD+0dA+dsGFP,SD+0dA+dsCBP-F1,SD+0dA+dsCBP-F2. Files in sft format can be opened with ScopePhoto. The ScopePhoto software can be downloaded from the address at the end of this document. * `fig1i_ovary size.csv` * `group` for different group include SD+0dA+dsGFP,SD+0dA+dsCBP-F1,SD+0dA+dsCBP-F2 * `figure` ovary figure name in fig1h_SD+dsCBP_ovary_phenotype.zip * `Length/Pixels` The pixel values of ovarian length measured using ScopePhoto * `1mm Scale/Pixels` The pixel values of 1mm Scale * `Actual Length` The actual length obtained by dividing Length/Pixels by 1mm Scale/Pixels * `Width/Pixels` The pixel values of ovarian width measured using ScopePhoto * `Actual Width` The actual Width obtained by dividing Width/Pixels by 1mm Scale/Pixels * `fig1j_SD+dsCBP_vg1_vg2_RTqPCR.CSV` * `group` for different group include 4-day-old female adults fat bodies of SD+dsGFP,SD+dsCBP * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of vg1` for ct value of vg1 * `ct value of vg2` for ct value of vg2 * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation * `fig1k_SD+dsCBP_JHAMT1_HMGR2_FPPS1_RTqPCR.CSV` * `group` for different group include 4-day-old female adults heads of SD+dsGFP,SD+dsCBP * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of JHAMT1` for ct value of JHAMT1 * `ct value of HMGR2` for ct value of HMGR2 * `ct value of FPPS1` for ct value of FPPS1 * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation * `fig1l_SD+dsCBP_JHAMT1_WB.jpg` include WB data of JHAMT1 in 2-day-old female adults heads of SD+dsGFP and SD+dsCBP * `fig1m_SD+dsCBP_JHAMT1_relative_protein_level.CSV` * `group` for different group include -day-old female adults heads SD+dsGFP1,SD+dsGFP2,SD+dsGFP3,SD+dsCBP1,SD+dsCBP2,SD+dsCBP3 * `JHAMT1` values of JHAMT1 protein * `Average value of JHAMT1 in GFP` average value calculation of JHAMT1 protein value * `Relative JHAMT1` caculation of relative JHAMT1 protein level * `α Tublin` values of α Tublin protein * `JAverage value of α Tublin in GFP` average value calculation of α Tublin protein value * `Relative α Tublin` caculation of relative α Tublin protein level * `JHAMT1/α Tublin` caculation of relative JHAMT1 protein level to α Tublin protein level * `fig1n_SD+dsCBP_HPLC_MSMS.CSV` * `Hemolymph volume` hemolymph volume from 4-day-old female adults of dsGFP and dsCBP is 50μL * `Concentrated volume` volume of sample after extraction, concentration, and reconstitution * `Concentration factor` concentration factor (fold) of the sample during preparation of the final assay solution * `JH III (ng/mL)=(Peak area+379686)/25553/Concentration factor` JH titer Calculation Formula * `group` for different group include SD+dsGFP1,SD+dsGFP2,SD+dsGFP3,SD+dsCBP1,SD+dsCBP2,SD+dsCBP3 * `Peak area` average value calculation of α Tublin protein value * `JH III (ng/mL)` caculation of relative α Tublin protein level * `fig1n_SD+dsCBP_HPLC_MSMS.jpg` include HPLC/MSMS data of standard JHIII and samples of SD+dsGFP and SD+dsCBP * `fig1o_SD+dsCBP_krh1_JHE1_RTqPCR.CSV` * `group` for different group include 4-day-old reproductive female adults fat bodies of SD+dsGFP,SD+dsCBP * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of krh1` for ct value of kr-h1 * `ct value of JHE1` for ct value of JHE1 * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation * `(fig2)` * `fig2a_SD+dsCBP_Nile_red.jpg` include nile red data of 4-day-old reproductive female adults fat bodies of SD+dsGFP and SD+dsCBP * `fig2b_SD+dsCBP_Nile_red_measurement.CSV` * `SD+dsGFP` group of SD+dsGFP include 40 lipid droplets * `SD+dsCBP` group of SD+dsCBP include 40 lipid droplets * `scale/μm` photo scale is 100μm * `pixel` pixel value corresponding to the scale bar * `length 1` measurement 1 * `length 2` measurement 2 * `average` average of the two measurements * `actual length/μm` caculation of actual length * `fig2b_SD+dsCBP_Nile_red_measurement.jpg` include nile red measurement pic * `fig2c_SD+dsCBP_TG.CSV` * `Group` group of Ethanol(blank),1/3 standard,standard,4-day-old reproductive female adults whole bodies of SD-dsGFP-4d-1,SD-dsGFP-4d-2,SD-dsGFP-4d-3,SD-dsGFP-4d-4,SD-dsGFP-4d-5,SD-dsCBP-4d-1,SD-dsCBP-4d-2,SD-dsCBP-4d-3,SD-dsCBP-4d-4,SD-dsCBP-4d-5 * `OD510` OD510 value * `average OD` average OD510 * `sample OD-blank OD` sample OD-blank OD * `standard OD-blank OD` standard OD-blank OD * `(sample OD-blank OD)/(standard OD-blank OD)` (sample OD-blank OD)/(standard OD-blank OD) * `Standard concentration(mmol/L)` Standard concentration(mmol/L) * `[(sample OD-blank OD)/(standard OD-blank OD)]*Standard concentration ` [(sample OD-blank OD)/(standard OD-blank OD)]*Standard concentration * `Fresh weight(gFw)` Fresh weight value of each sample * `sample TG concentration(gFW/L)` sample TG concentration(gFW/L) * `TG(mmol/gFw)` TG value * `average of SD-dsGFP` average TG value * `Relative TG=TG/(Average of SD-dsGFP)` relative TG Calculation Formula * `fig2d_SD+dsCBP_Total_lipid.CSV` * `Group` group of 4-day-old reproductive female adults whole bodies of SD+dsGFP-1 to SD+dsGFP-10,SD+dsCBP-1 to SD+dsCBP-10 * `Weight of the glass weighing bottle(W/g)` Weight of the glass weighing bottle(W/g) * `initial dry weight(IDW/g)` initial dry weight(IDW/g) * `dry weight after extration(EDW/g)` dry weight after extration(EDW/g) * `TL%=(IDW-EDW)/(IDW-W)×100 %` TL Calculation Formula * `fig2e_SD+dsCBP_HSP21_23_70_RTqPCR.CSV` * `group` for different group include 4-day-old reproductive female adults whole bodies of SD+dsGFP,SD+dsCBP * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of HSP21` for ct value of HSP21 * `ct value of HSP23` for ct value of HSP23 * `ct value of HSP70` for ct value of HSP70 * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation * `fig2f_SD+dsCBP_DP1_RTqPCR.CSV` * `group` for different group include 4-day-old reproductive female adults fat bodies of SD+dsGFP,SD+dsCBP * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of DP1` for ct value of DP1 * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation * `(fig3)` * `fig3aSDi+ME_ovary_phenotype.zip` include ovary phenotype pics and ovary size sft format files of 4-day-old reproductive female adults of SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+ME. Files in sft format can be opened with ScopePhoto. The ScopePhoto software can be downloaded from the address at the end of this document. * `fig3b_ovary_class.csv` * `group` for different group include SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+ME * `class` for different developmental class of ovaries include class 0,Class I,Class II,Class III,Class IV * `Standard` The criteria for classifying ovarian stages. Undifferentiated ovarian tubules lacking vitellogenin deposition=class 0,Ovarian tubule enlargement without vitellogenin deposition=Class I,Expanded ovarian tubules containing vitellogenin deposits but lacking defined eggs=Class II,Developed ovarian tubules with vitellogenin deposits and distinct immature eggs (spherical, size-variable precursors)=Class III,Fully matured ovarian tubules packed with uniform oval-shaped eggs=Class IV * `numbers` numbers of ovaries of class 0,Class I,Class II,Class III and Class IV * `%` Percentage of class 0,Class I,Class II,Class III oand Class IV * `fig3c_ovary_size.csv` * `group` for different group include SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+ME * `figure` ovary figure name in fig1h_SD+dsCBP_ovary_phenotype.zip * `Length/Pixels` The pixel values of ovarian length measured using ScopePhoto * `1mm Scale/Pixels` The pixel values of 1mm Scale * `Actual Length` The actual length obtained by dividing Length/Pixels by 1mm Scale/Pixels * `Width/Pixels` The pixel values of ovarian width measured using ScopePhoto * `Actual Width` The actual Width obtained by dividing Width/Pixels by 1mm Scale/Pixels * `fig3d_SDi+ME_RNAi_efficiency.CSV` * `group` for different group include 4-day-old reproductive female adults whole bodies of SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+ME * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of CBP` for ct value of CBP * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation * `fig3e_SDi+ME_JHE1_krh1_RTqPCR.CSV` * `group` for different group include 4-day-old reproductive female adults fat bodies of SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+ME * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of krh1` for ct value of kr-h1 * `ct value of JHE1` for ct value of JHE1 * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation * `fig3f_SDi+ME_vg1_vg2_RTqPCR.CSV` * `group` for different group include 4-day-old reproductive female adults fat bodies of SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+ME * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of vg1` for ct value of vg1 * `ct value of vg2` for ct value of vg2 * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation * `fig3g_SDi+ME_Nile_red.jpg` include nile red data of fat bodies of 4-day-old reproductive female adults of SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+ME * `fig3h_SDi+ME_Nile_red_measurement.CSV` * `SD+dsGFP+Ace` group of 4-day-old reproductive female adults fat bodies of SD+dsGFP+Ace include 40 lipid droplets * `SD+dsCBP+Ace` group of 4-day-old reproductive female adults fat bodies of SD+dsCBP+Ace include 40 lipid droplets * `SD+dsCBP+ME` group of 4-day-old reproductive female adults fat bodies of SD+dsCBP+ME include 40 lipid droplets * `scale/μm` photo scale is 100μm * `pixel` pixel value corresponding to the scale bar * `length 1` measurement 1 * `length 2` measurement 2 * `average` average of the two measurements * `actual length/μm` caculation of actual length * `fig3h_SDi+ME_Nile_red_measurement.jpg` include nile red measurement pic * `fig3i_SDi+ME_TG.CSV` * `Group` group of Ethanol(blank),1/3 standard,standard,4-day-old reproductive female adults whole bodies of SD-dsGFP-Ace-1,SSD-dsGFP-Ace-2,SD-dsGFP-Ace-3,SD-dsCBP-Ace-1,SD-dsCBP-Ace-2,SD-dsCBP-Ace-3,SD-dsCBP-ME-1,SD-dsCBP-ME-2,SD-dsCBP-ME-3 * `average OD` average OD510 * `sample OD-blank OD` sample OD-blank OD * `standard OD-blank OD` standard OD-blank OD * `(sample OD-blank OD)/(standard OD-blank OD)` (sample OD-blank OD)/(standard OD-blank OD) * `Standard concentration(mmol/L)` Standard concentration(mmol/L) * `[(sample OD-blank OD)/(standard OD-blank OD)]*Standard concentration ` [(sample OD-blank OD)/(standard OD-blank OD)]*Standard concentration * `Fresh weight(gFw)` Fresh weight value of each sample * `sample TG concentration(gFW/L)` sample TG concentration(gFW/L) * `TG(mmol/gFw)` TG value * `average of SD-dsGFP-Ace` average TG value * `Relative TG=TG/(Average of SD-dsGFP-Ace)` relative TG Calculation Formula * `fig3j_SDi+ME_total_lipid.CSV` * `Group` group of 4-day-old reproductive female adults whole bodies of SD+dsGFP+Ace-1 to SD+dsGFP+Ace-13,SD+dsCBP+Ace-1 to SD+dsCBP+Ace-13,SD+dsCBP+ME-1 to SD+dsCBP+ME-13 * `Weight of the glass weighing bottle(W/g)` Weight of the glass weighing bottle(W/g) * `initial dry weight(IDW/g)` initial dry weight(IDW/g) * `dry weight after extration(EDW/g)` dry weight after extration(EDW/g) * `TL%=(IDW-EDW)/(IDW-W)×100 %` TL Calculation Formula * `(fig4)` * `fig4a_LDi+ME_ovary_phenotype.zip` include ovary phenotype pics and ovary size sft format files of 4-day-old diapause female adults of LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+ME. Files in sft format can be opened with ScopePhoto. The ScopePhoto software can be downloaded from the address at the end of this document. * `fig4b_ovary_class.csv` * `group` for different group include LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+ME * `class` for different developmental class of ovaries include class 0,Class I,Class II,Class III,Class IV * `Standard` The criteria for classifying ovarian stages. Undifferentiated ovarian tubules lacking vitellogenin deposition=class 0,Ovarian tubule enlargement without vitellogenin deposition=Class I,Expanded ovarian tubules containing vitellogenin deposits but lacking defined eggs=Class II,Developed ovarian tubules with vitellogenin deposits and distinct immature eggs (spherical, size-variable precursors)=Class III,Fully matured ovarian tubules packed with uniform oval-shaped eggs=Class IV * `numbers` numbers of ovaries of class 0,Class I,Class II,Class III and Class IV * `%` Percentage of class 0,Class I,Class II,Class III oand Class IV * `fig4c_ovary_size.csv` * `group` for different group include LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+ME * `figure` ovary figure name in fig1h_SD+dsCBP_ovary_phenotype.zip * `Length/Pixels` The pixel values of ovarian length measured using ScopePhoto * `1mm Scale/Pixels` The pixel values of 1mm Scale * `Actual Length` The actual length obtained by dividing Length/Pixels by 1mm Scale/Pixels * `Width/Pixels` The pixel values of ovarian width measured using ScopePhoto * `Actual Width` The actual Width obtained by dividing Width/Pixels by 1mm Scale/Pixels * `fig4d_LDi+ME RNAi_efficiency_RTqPCR.CSV` * `group` for different group include 4-day-old reproductive female adults whole bodies of LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+ME * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of CBP` for ct value of CBP * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation * `fig4e_LDi+ME_Vg1_Vg2_RTqPCR.CSV` * `group` for different group include 4-day-old reproductive female adults fat bodies of LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+ME * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of vg1` for ct value of vg1 * `ct value of vg2` for ct value of vg2 * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation * `fig4f_LDi+ME_krh1_JHE1_RTqPCR.CSV` * `group` for different group include 4-day-old reproductive female adults fat bodies of LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+ME * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of krh1` for ct value of kr-h1 * `ct value of JHE1` for ct value of JHE1 * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation * `fig4g_LDi+ME_Nile_red.jpg` include nile red data of fat bodies of 4-day-old reproductive female adults of LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+ME * `fig4h_LDi+ME_Nile_Red_measurement.CSV` * `LD+dsGFP+Ace` group of LD+dsGFP+Ace include 40 lipid droplets * `LD+dsGFP+ME` group of LD+dsGFP+ME include 40 lipid droplets * `LD+dsCBP+Ace` group of LD+dsCBP+Ace include 40 lipid droplets * `LD+dsCBP+ME` group of LD+dsCBP+ME include 40 lipid droplets * `scale/μm` photo scale is 100μm * `pixel` pixel value corresponding to the scale bar * `length 1` measurement 1 * `length 2` measurement 2 * `average` average of the two measurements * `actual length/μm` caculation of actual length * `fig4h_LDi+ME_Nile_Red_measurement.jpg` include nile red measurement pic * `fig4i_LDi+ME_TG.CSV` * `Group` group of Ethanol(blank),1/3 standard,standard,4-day-old reproductive female adults whole bodies of LD-dsGFP-Ace-1,LD-dsGFP-Ace-2,LD-dsGFP-Ace-3,LD-dsGFP-Ace-4,LD-dsGFP-ME-1,LD-dsGFP-ME-2,LD-dsGFP-ME-3,LD-dsGFP-ME-4,LD-dsCBP-Ace-1,LD-dsCBP-Ace-2,LD-dsCBP-Ace-3,LD-dsCBP-Ace-4,LD-dsCBP-ME-1,LD-dsCBP-ME-2,LD-dsCBP-ME-3,LD-dsCBP-ME-4 * `average OD` average OD510 * `average OD` average OD510 * `sample OD-blank OD` sample OD-blank OD * `standard OD-blank OD` standard OD-blank OD * `(sample OD-blank OD)/(standard OD-blank OD)` (sample OD-blank OD)/(standard OD-blank OD) * `Standard concentration(mmol/L)` Standard concentration(mmol/L) * `[(sample OD-blank OD)/(standard OD-blank OD)]*Standard concentration ` [(sample OD-blank OD)/(standard OD-blank OD)]*Standard concentration * `Fresh weight(gFw)` Fresh weight value of each sample * `sample TG concentration(gFW/L)` sample TG concentration(gFW/L) * `TG(mmol/gFw)` TG value * `average of LD+dsGFP+Ace` average TG value * `Relative TG=TG/(Average of LD+dsGFP+Ace)` relative TG Calculation Formula * `fig4j_LDi+ME_total_lipid.CSV` * `Group` group of 4-day-old reproductive female adults whole bodies of LD+dsGFP+Ace-1 to LD+dsGFP+Ace-13,LD+dsGFP+ME-1 to LD+dsGFP+ME-13,LD+dsCBP+Ace-1 to LD+dsCBP+Ace-13,LD+dsCBP+ME-1 to LD+dsCBP+ME-13 * `Weight of the glass weighing bottle(W/g)` Weight of the glass weighing bottle(W/g) * `initial dry weight(IDW/g)` initial dry weight(IDW/g) * `dry weight after extration(EDW/g)` dry weight after extration(EDW/g) * `TL%=(IDW-EDW)/(IDW-W)×100 %` TL Calculation Formula * `(fig5)` * `fig5a_SDi+ME_TGL1_RTqPCR.CSV` * `group` for different group include 4-day-old reproductive female adults fat bodies of SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+ME * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of TGL1` for ct value of TGL1 * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation * `fig5b_LDi+ME_TGL1_RTqPCR.CSV` * `group` for different group include 4-day-old diapause female adults fat bodies of LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+ME * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of TGL1` for ct value of TGL1 * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation * `fig5c_SDi+ME_krh1_RTqPCR.CSV` * `group` for different group include 4-day-old reproductive female adults fat bodies of SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+ME * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of krh1` for ct value of Kr-h1 * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation * `fig5d_SDi+ME_Mcm4_RTqPCR.CSV` * `group` for different group include 4-day-old reproductive female adults fat bodies of SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+ME * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of Mcm4` for ct value of Mcm4 * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation * `fig5e_LDi+ME_krh1_RTqPCR.CSV` * `group` for different group include 4-day-old diapause female adults fat bodies of LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+ME * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of krh1` for ct value of Kr-h1 * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation * `fig5f_LDi+ME_Mcm4_RTqPCR.CSV` * `group` for different group include 4-day-old diapause female adults fat bodies of LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+ME * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of Mcm4` for ct value of Mcm4 * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation * `fig5g_SDi+ME_VgR_RTqPCR.CSV` * `group` for different group include 4-day-old reproductive female adults fat bodies of SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+ME * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of VgR` for ct value of VgR * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation * `fig5h_LDi+ME_VgR_RTqPCR.CSV` * `group` for different group include 4-day-old diapause female adults fat bodies of LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+ME * `biological replicate` for different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3 * `technical replicate` for different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2 * `ct value of VgR` for ct value of VgR * `ct value of rpl19` for ct value of rpl19 * `ct value of act1` for ct value of act1 * `Geometric mean of references` for mean ct values calculation * `ΔCt (target-reference)` for ΔCt values calculation * `mean ΔCt in technical replicate` for mean ΔCt in technical replicate calculation * `mean ΔCt in control group` for mean ΔCt in control group calculation * `ΔΔCt(target gene-reference)` for ΔΔCt(target gene-reference) calculation * `2^-ΔΔCt)` for 2^-ΔΔCt calculation **Description:** The data has been organized in the order of the figures. ## Code/software Scopephoto was used to measure the size of the ovaries or lipid droplets. [https://scopephoto.software.informer.com/3.1/](https://scopephoto.software.informer.com/3.1/)