DOI: 10.5061/dryad.tqjq2bw71
title: Data from: Essential function of transmembrane transcription factor MYRF in promoting transcription of miRNA lin-4 during C. elegans development

description: Precise developmental timing control is essential for organism formation
 and function, but its mechanisms are unclear. In C. elegans, the
 microRNA lin-4 critically regulates developmental timing
 by post-transcriptionally downregulating the larval-stage-fate controller
 LIN-14. However, the mechanisms triggering the activation
 of lin-4 expression toward the end of the first larval
 stage remain unknown. We demonstrate that the transmembrane transcription
 factor MYRF-1 is necessary for lin-4 activation. MYRF-1
 is initially localized on the cell membrane, and its increased cleavage
 and nuclear accumulation coincide with lin-4 expression
 timing. MYRF-1 regulates lin-4 expression
 cell-autonomously and hyperactive MYRF-1 can prematurely
 drive lin-4 expression in embryos and young first-stage
 larvae. The tandem lin-4 promoter DNA recruits
 MYRF-1GFP to form visible loci in the nucleus, suggesting that
 MYRF-1 directly binds to the lin-4 promoter. Our
 findings identify a crucial link in understanding developmental timing
 regulation and establish MYRF-1 as a key regulator of lin-4
 expression.
description: # Data from: Essential function of transmembrane transcription factor MYRF
 in promoting transcription of miRNA lin-4 during C. elegans development
 [https://doi.org/10.5061/dryad.tqjq2bw71](https://doi.org/10.5061/dryad.tqjq2bw71) The dataset comprises original images in CZI, TIF, or VSI formats, and datasheets in XLSX or XLS formats from quantitative and statistical analysis. The folder and file names typically indicate essential genotype and animal stage information. The files may be supplemented by additional notes in TXT format. CZI files were acquired using a Zeiss LSM880 microscope with Airyscan. VSI or TIF files were acquired using an Olympus BX63 microscope. Excel files (XLSX/XLS) are used for organizing and presenting data summaries and results of quantitative analyses. ## Description of the data and file structure Figure 1A: The dataset consists of a collection of representative images illustrating the subcellular localization patterns of GFP::MYRF-1 and the expression of the lin-4 transcriptional reporter (Plin-4-gfp) at different developmental stages. The first subfolder contains source images capturing animals exhibiting specific patterns of GFP::MYRF-1 subcellular localization during early-mid L1 (6 post-hatch hours), late L1 (15 post-hatch hours), and early L2 (20 post-hatch hours). The second subfolder includes source images depicting the expression of Plin-4-gfp in animals at distinct developmental stages, namely early-mid L1 (6 post-hatch hours), late L1 (15 post-hatch hours), and early L2 (20 post-hatch hours). Figure 1B: The folder contains raw data images of animals exhibiting a specific pattern of GFP::MYRF-1 subcellular localization at different developmental stages (early-mid L1 [6 post-hatch hours], late L1 [14 post-hatch hours], early L2 [20 post-hatch hours]) in .vsi files, along with the quantification of the GFP::MYRF-1 expression pattern in an .xlsx file. Figure 1C: The folder contains raw data images illustrating the expression of the lin-4 transcriptional reporter (Plin-4-gfp) in animals at distinct developmental stages, including early-mid L1 (6 post-hatch hours), late L1 (14 post-hatch hours), and early L2 (20 post-hatch hours), stored as .vsi files. Additionally, it includes quantification of the whole body Plin-4-gfp fluorescence intensity presented in an .xlsx file. Figure 2A: The raw data images illustrating the expression of Plin-4-gfp, a lin-4 transcriptional reporter, in wild-type (WT) and myrf-1(ju1121) animals at distinct developmental stages including early-mid L1 (6 post-hatch hours), late L1 (16 post-hatch hours), and early L2 (21 post-hatch hours). Figure 2B: This folder contains the raw data images illustrating the expression of the lin-4 transcriptional reporter (Plin-4-gfp) in wild-type (WT) and myrf-1(ju1121) animals at distinct developmental stages, including early-mid L1 (6 post-hatch hours), late L1 (16 post-hatch hours), and early L2 (21 post-hatch hours). These images are stored as .vsi files, accompanied by an .xlsx file containing quantification of the whole body Plin-4-gfp fluorescence intensity. Figure 2C: The raw data images illustrating the expression of the endogenous lin-4 transcriptional reporter (lin-4p::nls::mScarlet(umn84)) in wild-type (WT), myrf-1(ybq6), myrf-2(ybq42) mutants, and myrf-1; myrf-2 double mutants at early L2 stage (21 hours post-hatching). Figure 2D: This folder contains the raw data images illustrating the expression of the endogenous lin-4 transcriptional reporter (lin-4p::nls::mScarlet(umn84)) in wild-type (WT), myrf-1(ybq6), myrf-2(ybq42) mutants, and myrf-1; myrf-2 double mutants at early L2 stage (21 post-hatch hours) in .vsi format, along with the quantification of whole-body lin-4p::nls::mScarlet(umn84) fluorescence intensity provided in an .xlsx file. Figure 2E: The raw data images illustrating the expression of the endogenous lin-4 transcriptional reporter (lin-4p::nls::mScarlet(umn84)) in wild-type (WT) and myrf-1(ju1121) mutants at distinct developmental stages, including early-mid L1 (6 post-hatch hours), late L1 (14 post-hatch hours), and early L2 (21 post-hatch hours). Figure 2.F,Figure 2 S3 D: This folder contains the raw data images illustrating the expression of the endogenous lin-4 transcriptional reporter (lin-4p::nls::mScarlet(umn84)) in wild-type (WT) and myrf-1(ju1121) mutants at distinct developmental stages, including early-mid L1 (6 post-hatch hours), late L1 (14 post-hatch hours), and early L2 (21 post-hatch hours). The images are stored in .vsi files, while the quantification of whole body lin-4p::nls::mScarlet(umn84) fluorescence intensity is provided in an .xlsx file. Figure 2G: The raw data images depicting the expression of the endogenous lin-4 transcriptional reporter (lin-4p::nls::mScarlet(umn84)) in wild-type (WT) and pan-1(gk142) mutants at early L2 (21 post-hatch hours). Figure 2H: The raw data images depicting the expression of the endogenous lin-4 transcriptional reporter (lin-4p::nls::mScarlet(umn84)) in wild-type (WT) and pan-1(gk142) mutants at early L2 (21 post-hatch hours).The images are stored in .vsi files, while the quantification of whole body lin-4p::nls::mScarlet(umn84) fluorescence intensity is provided in an .xlsx file. Figure 2 S1.A: This folder contains the raw data images depicting the expression of the lin-4 transcriptional reporter (Plin-4-gfp) in wild-type (WT), myrf-1(syb1491) and myrf-1(syb1468) mutants at L2 stage (24 post-hatch hours)). Figure 2 S1.B: The folder contains .vsi files of raw data images illustrating the expression of the lin-4 transcriptional reporter (Plin-4-gfp) in wild-type (WT), myrf-1(syb1491), and myrf-1(syb1468) mutants at L2 stage (24 post-hatch hours). Additionally, an .xlsx file provides quantification of the whole body Plin-4-gfp fluorescence intensity. Figure 2 S2: The raw data images illustrating the expression of the endogenous lin-4 transcriptional reporter (lin-4p::nls::mScarlet(umn84)) in umn84 homozygous and heterozygous animals at 6, 14, 21,33 and 45 hours post-hatch. Figure 2 S3.A: The raw data images depicting the expression of the endogenous lin-4 transcriptional reporter (lin-4p::nls::mScarlet(umn84)) in myrf-1(ybq6)/+, myrf-1(ybq6), and myrf-1; myrf-2 double mutants at early L2 (21 post-hatch hours). Figure 2 S3.B: The raw data images depicting the expression of the endogenous lin-4 transcriptional reporter (lin-4p::nls::mScarlet(umn84)) in wild-type (WT) and pan-1(gk142) mutants at early L2 (21 post-hatch hours). Figure 2 S3.C: The raw data images depicting the expression of the endogenous lin-4 transcriptional reporter (lin-4p::nls::mScarlet(umn84)) in wild-type (WT) and myrf-1(ju1121) mutants at early L2 (21 post-hatch hours). Figure 2 S4.A: The images illustrating the expression of the lin-4 transcriptional reporter (Plin-4-gfp) in L2, L3, and L4 animals were treated with or without 4 mM K-NAA for a duration of 12 hours. Figure 2 S4.B: The images illustrating the expression of the lin-4 transcriptional reporter (Plin-4-gfp) in L2, L3, and L4 animals were subjected to treatment with or without 4 mM K-NAA for a duration of 12 hours. Additionally, it includes quantification of the whole body Plin-4-gfp fluorescence intensity, presented as .xlsx files. Figure 3A: The raw data images depicting the expression of the endogenous LIN-14::GFP(cc2841) in wild-type and myrf-1(ju1121) at the early L1 (0h), late L1 (16h), and early L2 (21h) stages. Figure 3B: The folder contains raw data images illustrating the expression of endogenous LIN-14::GFP(cc2841) in wild-type and myrf-1(ju1121) at early L1 (0h), late L1 (16h), and early L2 (21h) stages, saved as .vsi files. Additionally, an .xlsx file provides quantification of the head LIN-14::GFP fluorescence intensity. Figure 4A: The folder contains raw data images illustrating the expression of the lin-4 transcriptional reporter (Plin-4-gfp) in wild-type (WT), myrf-1(ju1121) mutants, and myrf-1(ju1121) mutants carrying transgenes Pmyo-3-myrf-1 and Pdpy-7-myrf-1, respectively. Figure 4B: The images depict the expression of the lin-4 transcriptional reporter (Plin-4-gfp) in animals with tissue-specific ablation of myrf-1 in the epidermis (myrf-1LoxP(ybq98), Pdpy-7-NLS::Cre(tmIs1028)) and control (myrf-1LoxP(ybq98)) animals. Figure 4C: This folder contains the raw data images depicting the lin-4 transcriptional reporter (Plin-4-gfp) in animals with tissue-specific ablation of myrf-1 in the epidermis (myrf-1LoxP(ybq98), Pdpy-7-NLS::Cre(tmIs1028)) and control (myrf-1LoxP(ybq98)) animals at L2(24h) in .vsi files, as well as the quantification of Plin-4-gfp fluorescence intensity presented in an .xlsx file. Figure 4E: The bright field images of animals with tissue-specific ablation of myrf-1 in the epidermis (myrf-1LoxP(ybq98), Pdpy-7-NLS::Cre(tmIs1028)) and control (myrf-1LoxP(ybq98)) animals were captured at Day 1 and Day 2 adult stages, corresponding to 72 hours and 84 hours post-hatching, respectively. Figure 5A: The images depicting the expression of the endogenous lin-4 transcriptional reporter (lin-4p::nls::mScarlet(umn84)) at embryos ,early L1 and late L1 stage. Figure 5B: The images show the expression of the endogenous lin-4 transcriptional reporter (lin-4p::nls::mScarlet(umn84)) in animals with the Prpl-28-GFP::myrf-1(Δ601-650) transgene and in control animals without the transgene, at the embryo and early L1 stages. Figure 5C: This folder contains raw data images showing the expression of the lin-4 transcriptional reporter (Plin-4-gfp) in wild-type (WT) and myrf-1(syb1313) animals at different developmental stages: early-mid L1 (6 hours post-hatch) and late L1 (15 hours post-hatch). Figure 5D: The folder contains raw data images illustrating the expression of the lin-4 transcriptional reporter (Plin-4-gfp) in wild-type (WT) and myrf-1(syb1313) animals at early-mid L1 (6 post-hatch hours) and late L1 (15 post-hatch hours), stored as .czi files. Additionally, it includes quantification of Plin-4-gfp fluorescence intensity in neurons and pharynx, presented as .xlsx files. Figure 5E: This folder contains raw data images depicting the expression of the endogenous lin-4 transcriptional reporter (lin-4p::nls::mScarlet(umn84)) in wild-type (WT) and myrf-1(syb1313) animals at distinct developmental stages: early-mid L1 (6 hours post-hatch), late L1 (14 hours post-hatch), and early L2 (21 hours post-hatch). Figure 5F: The folder contains raw data images illustrating the expression of the endogenous lin-4 transcriptional reporter (lin-4p::nls::mScarlet(umn84)) in wild-type (WT) and myrf-1(syb1313) animals at distinct developmental stages (early-mid L1 (6 post-hatch hours), late L1 (14 post-hatch hours) and early L2(21h). Additionally, it includes quantification of lin-4p::nls::mScarlet fluorescence intensity in whole body, neurons and pharynx, presented as .xlsx files. Figure 6D: The bright field images of wild-type (WT) and transgenic animals carrying the Plin-4-gfp (maIs134) transgene were captured at the L2 stage (26 hours post-hatch). Figure 6E: The bright field images of wild-type (WT) and transgenic animals carrying the Plin-4-gfp (maIs134) transgene were captured at 14, 19, 22, 26, 30, 36, 42, and 48 hours post-hatch. Figure 6F,G: The folder contains raw data images showing the expression of GFP::MYRF-1 (ybq14) and TetR::tagRFP (ybqSi233) in animals with lin-4 promoter (2.4 kb)-7xTetO sequence DNA arrays, 7xTetO sequence-containing DNA arrays, and control animals without DNA arrays respectively. Figure 6I: The folder contains raw data images showing the expression of GFP::MYRF-1 (ybq14) and TetR::tagRFP (ybqSi233) in animals with lin-4 promoter (2.4 kb)-7xTetO sequence DNA arrays, 7xTetO sequence-containing DNA arrays, and control animals without DNA arrays respectively. Additionally, it includes quantification of GFP::MYRF-1 and TetR::tagRFP fluorescence intensity in bar ROIs centered at the fluorescent spot, presented as .xlsx files. Figure 6 S1.A: Bright field images of wild-type (WT) animals, transgenic animals carrying the Plin-4-gfp (maIs134) transgene, and animals carrying both the Plin-4-gfp (maIs134) and myrf-1LoxP (ybqIs112) transgenes were captured at 70 hours post-hatch. Figure 6 S1.B: The bright field images of wild type (WT), transgenic animals carrying the Plin-4-gfp (maIs134) transgene, and animals carrying both the Plin-4-gfp (maIs134) and myrf-1LoxP(ybqIs112) transgenes were captured at 48 hours post-hatch. Figure 6 S2.A,B,C: The folder contains raw data images illustrating the expression of GFP::MYRF-1(ybq14) in animals harboring both co-marker (Pmyo-2-mCherry) and lin-4 promoter (2.4 kb) DNA arrays, as well as animals carrying only the co-marker. Additionally, it includes quantification of GFP::MYRF-1 fluorescence intensity in bar ROIs, centered at the fluorescent spot, presented as .xlsx files. Figure 7C: This folder contains the raw data images depicting the expression of Pmir-48-gfp(ybqSi206), Pmir-73-gfp(ybqSi208), and Pmir-230-gfp(ybqSi209) transgene in wild-type and myrf-1(ju1121) animals at early-mid L1 (6h) and middle L2 (24h). Figure 7D: This folder contains the raw data images depicting the expression of Pmir-48-gfp(ybqSi206), Pmir-73-gfp(ybqSi208), and Pmir-230-gfp(ybqSi209) in wild-type and myrf-1(ju1121) animals at early-mid L1 (6h) and middle L2 (24h) in .vsi files. Additionally, it includes quantification of Pmir-48-gfp(ybqSi206), Pmir-73-gfp(ybqSi208), and Pmir-230-gfp(ybqSi209) fluorescence intensity, presented as .xlsx files.