EMBO/SourceData huggingface dataset:
Figure legend: "(B) RT-qPCR analyses of Hnf4a, Nr1h4 and Foxa2 expression in MPH treated with vehicle (Control) or 1µM thapsigargin (ERS) for 1h or 4h (4 independent experiments). The bar graph shows means ± SD (standard deviations). One-sample t-test with BH correction for multiple testing was used to determine if the mean Log2 FC ERS/Control is statistically different from 0, *P &lt; 0.05."
NCBI gene IDs:
"Foxa2: 15376
Hnf4a: 15378
Nr1h4: 20186"
Figure legend: "(B) Box plots showing Log2 FC ERS/Control in mouse liver for ERS UP genes 8h and 34h after tunicamycin injection in WT or ATF6 KO mice (3 mice per experimental condition). Box plots are composed of a box from the 25th to the 75th percentile with the median as a line and min to max as whiskers. One-sample t-test with BH correction for multiple testing was used to determine if the mean Log2 FC ERS/Control is statistically different from 0, *P &lt; 0.05. NS, not significant. (C) Similar analyses to panel (B) for ERS DOWN and LIVER-ID genes (3 mice per experimental condition). Box plots are composed of a box from the 25th to the 75th percentile with the median as a line and min to max as whiskers. One-sample t-test with BH correction for multiple testing was used to determine if the mean Log2 FC ERS/Control is statistically different from 0, *P &lt; 0.05. "
NCBI gene IDs:
"ATF6: 226641"
Figure legend: "(D) Main observations from (Huck et al., 2019) (left) and box plots showing Log2 FC HNF4A KO/Control in mouse liver (3 mice per group) for LIVER-ID genes and TFs 5 days after PHx (right). Box plots are composed of a box from the 25th to the 75th percentile with the median as a line and min to max as whiskers. One-sample t-test with BH correction for multiple testing was used to determine if the mean Log2 FC HNF4A KO/Control is statistically different from 0, *P &lt; 0.05."
NCBI gene IDs:
"HNF4A: 15378"
Figure legend: "F - Left: relative mRNA expression of Chat in ChATfl/fl and ChATfl/fl;Vav-iCre IWAT (n = 9). Chat expression was analyzed by qPCR and normalized to levels of Tbp using the 2-ΔΔCt method. Right: quantification by LC/MS-MS of Ach concentration in SVF isolated from ChATfl/fl and ChATfl/fl;Vav-iCre IWAT (n = 6)."
NCBI gene IDs:
"Chat: 12647
ChAT: 12647
iCre: 2777477
Tbp: 21374
Vav: 22324"
Figure legend: "Body weight (C) of ChATfl/fl;LysM-Cre (n = 7 for ChATfl/fl, n = 8 for Cre ChATfl/fl;Cd4-Cre (n = 6 for ChATfl/fl and n = 7 for Cre ChATfl/fl;Mb1-Cre (n = 12 in C, n = 9 for ChATfl/fl, n = 8 for Cre and littermate ChATfl/fl mice housed at RT."
NCBI gene IDs:
"Cd4: 12504
Mb1: 12518
ChAT: 12647
Cre: 2777477
LysM: 17105"
Figure legend: "E-G - mRNA expression of Chrna2 and thermogenic genes in IWAT of ChATfl/fl;LysM-Cre (n = 24 for ChATfl/fl_RT, n = 25 for Cre_RT, n = 20 for ChATfl/fl_CE, n = 20 for Cre_CE) (E), ChATfl/fl;Cd4-Cre (n = 17 for ChATfl/fl_RT, n = 22-23 for Cre_RT, n = 19 for ChATfl/fl_CE, n = 22 for Cre_CE) (F), ChATfl/fl;Mb1-Cre (n = 10 for ChATfl/fl_RT, n = 11 for Cre_RT, n = 9-12 for ChATfl/fl_CE, n = 13 for Cre_CE) (G) and littermate ChATfl/fl mice housed at RT or 4oC (CE) for 6 h. An insert graph in (E) highlights mRNA expression of Chrna2 and thermogenic genes in IWAT of ChATfl/fl and ChATfl/fl;LysM-Cre mice after 6 h CE."
NCBI gene IDs:
"Cd4: 12504
Mb1: 12518
ChAT: 12647
Chrna2: 110902
Cre: 2777477
LysM: 17105"
Figure legend: "K - Average whole-body oxygen consumption rate (OCR) of ChATfl/fl (n = 10) and ChATfl/fl;LysM-Cre (n = 14) mice housed in metabolic chambers at RT or CE for 6 h (from 9 a.m. to 3 p.m.)."
NCBI gene IDs:
"ChAT: 12647
Cre: 2777477
LysM: 17105"
Figure legend: "J - Relative mRNA expression of Adrb1, Adrb2 and Adrb3 in IWAT MΦ sorted from WT mice (n = 4). Expression was measured by qPCR and normalized to levels of Tbp using the 2-ΔΔCt method."
NCBI gene IDs:
"Adrb1: 11554
Adrb2: 11555
Adrb3: 11556
Tbp: 21374"
Figure legend: "F - LC/MS-MS quantification of acetylcholine levels secreted by SVF cells derived from IWAT of β2WT and β2KO mice treated with veh or 1 mg/kg Form for 2 h (n = 3)."
NCBI gene IDs:
"β2: 11555"
Figure legend: "M - Left: BMDMs were treated for 2 h with veh or pan β-AR agonist (100 μM NE), β2-AR antagonist (5 μM butoxamine, Buto) or a combination of NE and β2 antagonist (Buto) (n = 6). Chat mRNA expression was measured by qPCR and normalized to levels of Tbp using the 2-ΔΔCt method. Right: Total number of ChAT-eGFP+ BMDMs. ChAT-eGFP BMDMs were treated for 2 h with veh or pan β-AR agonist (100 μM NE), β2-AR antagonist (5 μM butoxamine, Buto) or a combination of NE and β2-AR antagonist (Buto) (n = 4). An equal number of events (50,000) were analyzed by flow cytometry."
NCBI gene IDs:
"Chat: 12647
Tbp: 21374"
Figure legend: "N - Left: Bicompartmental co-culture system with media alone (Ctrl) or WT SVF cells isolated from IWAT in the upper compartment (transwell insert) and freshly isolated IWAT explants from β-less mice in the lower compartment. Cells were co-cultured for 4 h in the presence or absence of β2-AR agonist (2.5 μM Form). 150 μM rivastigmine was added to the media to prevent degradation of Ach. Right: qPCR analyses of Chat and Ucp1 mRNA levels in β‑less explants following co-culture with media (n = 4), vehicle (n = 10) or β2-AR agonist (n = 10) treated SVF cells. mRNA expression was measured by qPCR and normalized to levels of Tbp using the 2-ΔΔCt method."
NCBI gene IDs:
"Chat: 12647
Tbp: 21374
Ucp1: 22227"
Figure legend: " C Relative mRNA expression of Apopt1 normalized to the expression of GAPDH in skeletal muscle and liver of wild-type, heterozygous and knock-out mice of three months of age. Data are presented as mean ± SEM (n = 6 mice per genotype). The asterisks represent the significance levels calculated by two-way ANOVA with Sidak's multiple comparisons test: Muscle - ***P = 0.0001 (WT vs KO), *P = 0.0310 (WT vs het), *P = 0.0395 (het vs KO), Liver - **P = 0.0022 (WT vs KO), *P = 0.0101 (WT vs het) "
NCBI gene IDs:
"Apopt1: 68020"
Figure legend: " A APOPT1HA was expressed in S6 and S2 immortalized fibroblasts as shown by the Western blot immunovisualization. The expression levels were tested at different days after transduction. The graph on the right shows the activities of CIV normalized to CS in three biological replicates per cell line. Data are presented as mean ± SEM (n = 3). The asterisks represent the significance levels calculated by two-way ANOVA with Tukey's multiple comparisons test: **P = 0.0030 (S6 naïve vs S6 APOPT1HA), *P = 0.0165 (S6 EV vs S6 APOPT1HA )"
NCBI gene IDs:
"APOPT1: 84334"
Figure legend: " B APOPT1 GFP was expressed in S6 and S2 immortalized fibroblasts as shown by the Western blot immunovisualization. The graph on the right shows the activities of CIV normalized to CS in four biological replicates per cell line. Data are presented as mean ± SEM (n = 4). The asterisks represent the significance levels calculated by two-way ANOVA with Tukey's multiple comparisons test: **P = 0.0066 (S6 naïve vs S6 APOPT1GFP), ***P = 0.0006 (S6 EV vs S6 APOPT1GFP), *P = 0.0169 (S2 naïve vs S2 APOPT1GFP), ***P = 0.0008 (S2 EV vs S2 APOPT1GFP)"
NCBI gene IDs:
"APOPT1: 84334"
Figure legend: " E L-[35S]-Methionine pulse-chase labeling of mtDNA-encoded proteins. After a two-hour exposure with the radioactive label (pulse), cells were cultured in cold medium for the indicated chase times. The graphs show the densitometric quantification of the bands corresponding to MT-CO1 (left graph) and MT-CO2+MT-CO3 (right graph) normalized to the ATP6 band over the indicated time points. Graphs represent the values of three biological replicas for each cell line. Data are presented as mean ± SEM (n = 4 for controls, n = 2 for S2/S6/S2 APOPT1GFP/S6 APOPT1GFP). The asterisks represent the significance levels calculated by two-way ANOVA with Tukey's multiple comparisons test: MT-CO1 - **P = 0.0039 ( 3.5 hours, controls vs S6), *P = 0.0202 (3.5 hours, controls vs S2), *P = 0.0163 (5 hours, S6vs S6 APOPT1GFP), *P = 0.0118 (5 hours, controls vs S2), **P = 0.0011 (20 hours, controls vs S6), *P = 0.0343 (20 hours, S2 vs S2 APOPT1GFP), ***P = 0.0007 (20 hours, S6 vs S6 APOPT1GFP ), ***P = 0.0002 (20 hours, controls vs S2), *****P &lt; 0.0001 (20 hours, controls vs S6), MT-CO2/MT-CO3 - *P = 0.0194 (3.5 hours, S6 vs S6 APOPTGFP), ***P = 0.0003 (3.5 hours, controls vs S6), *P = 0.0375 (5 hours, S6 vs S6 APOPT1GFP), *P = 0.0234 (20 hours, S2 vs S2 APOPT1GFP), ****P &lt; 0.0001 (20 hours, S6 vs S6 APOPT1GFP), **P = 0.0013 (20 hours, controls vs S2), ****P &lt; 0.0001 (20 hours, controls vs S6)"
NCBI gene IDs:
"APOPT: 84334
APOPT1: 84334"
Figure legend: " C Western blot analysis of SDS-PAGE of total lysates from 143B cells overexpressing tagged APOPT1 (as indicated) and exposed to 100 µM H2O2, as illustrated by the scheme (H2O2 treatment), for the indicated times. The graphs represent the densitometric quantification of the tagged APOPT1 signal at each time point. The graph inset shows that the increase of APOPT1 occurs in the first minutes after the exposure to H2O2 "
NCBI gene IDs:
"APOPT1: 84334"
Figure legend: " D Western blot analysis of SDS-PAGE of total lysates from 143B cells overexpressing tagged APOPT1 (as indicated) and exposed to 5 µM MitoParaquat (MitoPQ), as illustrated by the scheme (MitoPQ treatment), for the indicated times. The graphs represent the densitometric quantification of the tagged APOPT1 signal at each time point. The graph inset shows that the increase of APOPT1 occurs in the first minutes after the exposure to MitoPQ "
NCBI gene IDs:
"APOPT1: 84334"
Figure legend: " A Western blot analysis of SDS-PAGE of different mitochondrial proteins in total lysates from the indicated cell lines treated with 5 µM MitoPQ at the indicated times. UT: untreated cells. B Densitometric quantification of APOPT1GFP signal during the treatment in two biological replicas. C Densitometric quantification of MT-CO1 signal in the non-complemented APOPT1-less cells (S6 GFP) vs. the complemented cells (S6 APOPT1GFP). Three biological replicas (n=3) were carried out for each cell line. The signals in UT S6 APOPT1 were considered 100%. The levels of MT-CO1 were significantly lower in the S6 GFP patient cells after 20 hours of MitoPQ treatment compared to the untreated cells (*P = 0.0202, two-tailed unpaired Student's t-test). D Densitometric quantification of MT-CO2 signal in the non-complemented APOPT1-less cells (S6 GFP) vs. the complemented cells (S6 APOPT1GFP). Three biological replicas were carried out for each cell line. The signals in UT S6 APOPT1 were considered 100% "
NCBI gene IDs:
"APOPT1: 84334"
Figure legend: "C Co-IP with  HEK293T cells co-transfected with plasmids encoding   Hwa-Flag and ZNRF3-HA or ZNRF3ΔRING. Lysates  immunoprecipitated with anti-Flag were immunoblotted with HA  and Flag antibodies. Blots are representative of 3 independent experiments. D Co-IP  with HEK293T  cells co-transfected with plasmids encoding Hwa-Flag and ZNRF3-HA or ZNRF3ΔRING. Lysates  immunoprecipitated  with anti-HA were immunoblotted with  Flag and HA antibodies. Blots are representative of 3 independent experiments. "
NCBI gene IDs:
"Flag: 
HA: 
Hwa: 108718903
ZNRF3: 108717096"
Figure legend: "E Colocalization of Hwa-Flag (red) and  ZNRF3-HA (green) or ZNRF3ΔRING-HA (green) in Hela cells is indicated by yellow dots. Baf-A1, 50 nM, 12 hrs. Magnification: 100 ×; scale bar: 10 μm."
NCBI gene IDs:
"HA: 
ZNRF3: 108717096"
Figure legend: "B Whole mount  in situ hybridization of Spemann organizer markers  chrd, gsc, and xnr3 in gastrula embryos (stage 10.5). Oocytes were injected with AS or mRNAs as indicated, Dosage: znrf3 AS (3 ng AS4 + 3 ng AS9), znrf3  mRNA 100 pg, znrf3ΔRING mRNA 50 pg. embryos were obtained through Host transfer technology. Scale bars: 1 mm."
NCBI gene IDs:
"chrd: 108716188///108716939///398045
gsc: 397748///397752
xnr3: 373597///378537
znrf3: 108717096"
Figure legend: "H Whole mount in situ hybridization of Spemann organizer markers chrd and gsc in  gastrula embryos (stage 10.5). Embryos were injected and obtained as descripted in (G). Scale bars: 1 mm."
NCBI gene IDs:
"chrd: 108716188///108716939///398045
gsc: 397748///397752"
Figure legend: "I Representative phenotypes  of  tailbud stage Xenopus leavis embryos injected with 500 pg mRNAs of ctrl (gfp mRNA), znrf3FL, and znrf3 truncations as indicated at 4-cell stage. Duplicated axes (red arrowhead). znrf3FL, full-length znrf3; znrf3ΔRING, znrf3 lacking the RING domain; znrf31-320, amino acids  1-320 of  znrf3; znrf3320-606, amino acids 320-606 of znrf3; znrf3606-854, amino acids 606-854 of znrf3. Scale bars: 1 mm."
NCBI gene IDs:
"gfp: 
znrf3: 108717096"
Figure legend: "B Hwa ubiquitination was analyzed under denaturing conditions. HEK293T cells co-transfected with wildtype His-HA-Ubiquitin (His-HA-Ub WT) or lysine-less mutant (K0), Hwa-Flag, and  ZNRF3-Myc. The lysates were immunoblotted with Myc,  Flag, and α-tub (α-tubulin). Lysates immunoprecipitated  with anti-Flag were immunoblotted  with HA and Flag  antibodies. K0, all lysine sites on ubiquitin were mutated. Blots  are representative of 3 independent experiments."
NCBI gene IDs:
"Flag: 
HA: 
His: 
Myc: 
Hwa: 108718903
ZNRF3: 108717096"
Figure legend: "D Screen of ZNRF3-mediated Hwa ubiquitination  sites under denaturing conditions. HEK293T cells were co-transfected with His-HA-Ub, Hwa-Flag or  Hwa10KR-Flag, and  ZNRF3 or ZNRF3ΔRING. The lysate was  immunoblotted  with Myc, Flag, and α-tub (α-tubulin). Lysates immunoprecipitated with anti-Flag were  immunoblotted  with HA and Flag antibodies. Hwa10KR-Flag mutant, all lysine sites in Hwa  sequence were replaced with arginine. Denaturing conditions: before  immunoprecipitation, lysates were denatured by adding SDS to a final concentration at 1% and boiling at 95℃ for 15 min. Blots  are representative of 3 independent experiments."
NCBI gene IDs:
"Flag: 
HA: 
His: 
Hwa: 108718903
ZNRF3: 108717096"
Figure legend: "A Western blot  analysis of Hwa in HEK293T cells co-transfected with Hwa-Flag, and ZNRF3-HA   (5 μg) or ZNRF3ΔRING (2.5 μg). Lysate was immunoblotted with  Flag, HA, and GAPDH  antibodies.  Blots  are representative of 3 independent experiments. B Densitometry analysis of Hwa  bands from the western blot in (A) for corresponding lanes normalized to GAPDH. n = 3 independent experiments, mean ± SEM, ns p &gt; 0.05, **p &lt; 0.01 by unpaired t test. "
NCBI gene IDs:
"Flag: 
HA: 
Hwa: 108718903
ZNRF3: 108717096"
Figure legend: "C Western blot analysis of Hwa in HEK293T cells co-transfected with Hwa-Flag and ZNRF3-HA. Cells were treated with or without Baf-A1 (50 nM) or MG132 (20 μM) for about 12 hours. Lysates were immunoblotted with  Flag, HA, and  α-tub (α-tubulin) antibodies.  Blots are representative of 3 independent experiments. D Densitometry analysis of Hwa bands from the western blot in (C) for corresponding lanes normalized to α-tubulin. n = 3 independent experiments, mean ± SEM, ns p &gt; 0.05, **p &lt; 0.01, ****p &lt; 0.0001 by unpaired t test. "
NCBI gene IDs:
"Flag: 
HA: 
Hwa: 108718903
ZNRF3: 108717096"
Figure legend: "F Colocalization by immunofluorescence of  Hwa (green signals) with  lysosome (LAMP1, red signals) in Hela  cells. Cells were co-transfected with Hwa-Flag, and  ZNRF3-HA  or ZNRF3ΔRING-HA. Magnification, 100 ×; scale bars: 10 μm."
NCBI gene IDs:
"Flag: 
HA: 
Hwa: 108718903
ZNRF3: 108717096"
Figure legend: "I Surface biotinylation and streptavidin pulldown  of Hwa-Myc-Flag. HEK293T cells were co-transfected with Hwa-MF (Hwa-Myc-Flag), and ZNRF3-Myc or  ZNRF3ΔRING-Myc. Lysate was immunoblotted  with Myc, Flag, and GAPDH  antibodies. Lysates immunoprecipitated  with anti-Flag (2nd elution) were immunoblotted with Flag and HA  antibodies."
NCBI gene IDs:
"Flag: 
Myc: 
Hwa: 108718903
ZNRF3: 108717096"
Figure legend: "C Whole mount in situ hybridization of Spemann organizer markers chrd  and gsc in gastrula embryos. Embryos  were obtained as descripted in (B). Scale bars: 1mm."
NCBI gene IDs:
"chrd: 108716188///108716939///398045
gsc: 397748///397752"
Figure legend: "(D) NIH 3T3 cells transfected by equal amounts of plasmid fused with gfp (without ATG) to the sequence from 5' cap of pri-miR-31 to ATG site or which mutated to ATT of the sORF. Cells were selected by G418 for 2 days. Analysis of the GFP expression by confocal microscopy or flow cytometry, scale bar 100 μm."
NCBI gene IDs:
"gfp: 
pri-miR-31: 723895"
Figure legend: "(E) Immunoblot analysis of GFP and miPEP31 in NIH 3T3 cells transfected by pEGFP-N1-miPEP31 or pEGFP-N1, the red arrow indicate predicted size of miPEP31-GFP band."
NCBI gene IDs:
"EGFP: "
Figure legend: "(F) Flow-fish to detect the localization of pri-miR-31. The location of pri-miR-31 was analyzed by confocal microscopy. Scale bar 5 μm."
NCBI gene IDs:
"pri-miR-31: 723895"
Figure legend: "(H) Immunoblot analysis of spleen, lymph node and thymus from WT or miR-31-/- mice with miPEP31 antibody, the red arrow indicate predicted size of miPEP31."
NCBI gene IDs:
"miR-31: 723895"
Figure legend: "(B, D) Yeast cells of the indicated genotype were transformed with plasmids coding for Atg5 and Atg16, respectively, and subjected to fractionation experiments. The proteins were detected by anti‐Myc western blotting. The presence of the protein in the pellet fraction indicated membrane binding."
NCBI gene IDs:
"Atg16: 326115
Atg5: 855954"
Figure legend: "(C) Quantification showing the relative amounts of Atg5-9 × Myc and Atg5-9 × Myc-Atg12 in the pellet fractions relative to Pex30. The amount of Atg5-9 × Myc and Atg5-9 × Myc-Atg12 in atg5Δ cells was set to 1. See Supplementary Figure 1 for graph based on non‐normed data. The quantification is based on three independent experiments and the averages and the standard deviations are shown. The numbers next to the blots indicate the molecular weight in kDa. P, pellet; S, supernatant. Figure source data can be found with the Supplementary data."
NCBI gene IDs:
"Atg12: 852518
atg5: 855954"
Figure legend: "(C) Yeast cells were transformed with the indicated expression constructs and Atg5-2 × Myc was immunoprecipitated with an anti‐Myc antibody. The precipitated protein was subjected to anti‐Atg5 and anti‐Atg12western blotting. The substitution of R171 and K160 to glutamate caused the mutant protein to run higher. The same phenomenon was observed for the recombinant protein ( Supplementary Figure 3)."
NCBI gene IDs:
"Atg5: 855954"
Figure legend: "(E) Anti‐Myc and anti‐Pex30western blots of yeast cell fractions from cells of the indicated genotype and expressing Atg16 and Atg5 or Atg5 K160E, R171E."
NCBI gene IDs:
"Atg5: 855954"
Figure legend: "(A) Yeast cells of the indicated genotype expressing Atg5-3 × mCherry or Atg5 K160E, R171E-3 × mCherry were treated with rapamycin and imaged using a Deltavision microscope. Maximum projections of z‐stacks are shown."
NCBI gene IDs:
"Atg5: 855954"
Figure legend: "G The entry efficiency of SARS-CoV-2 and RshSTT182 pseudoviruses in HeLa-hACE2 cells. Green fluorescence indicates the entry of the pseudoviruses into the Hela-hACE2 cells. Unaltered HeLa cells were used as a negative control. H Statistics for transduction of pseudoviruses. Data represents the results of five replicates in one representative assay. All data are presented as mean ± SD. Pseudovirus infection assays were performed at least twice."
NCBI gene IDs:
"ACE2: 59272"
Figure legend: "B. RT-PCR analysis for forebrain (FOXG1), optic cup (VSX2), dorsal forebrain (PAX6 and EMX2) and ventral forebrain (NKX2-1) identity in organoids. All expression values (2-ΔCt) calculated relative to reference gene TBP. Analysis was performed on 5-6 organoids from multiple independent differentiations - control (7), dorsal (7), extended ventral (9) and short ventral (5). Significance values, * = &lt;0.05, ** = &lt;0.01, *** = &lt;0.001. Statistical analysis was performed using one-way ANOVA and post-hoc Tukey's comparison of means. The central band depicts the median, the boxes depict values between lower and upper quartile and the whiskers display the minimum and maximum values."
NCBI gene IDs:
"EMX2: 2018
FOXG1: 2290
NKX2-1: 7080
PAX6: 5080
TBP: 6908
VSX2: 338917"
Figure legend: "C. UMAPs depicting the expression of GABAergic markers DLX2 and DLX5, progenitor marker TOP2A, MGE marker SOX6, CGE marker NR2F2, and striatal marker ZFHX3."
NCBI gene IDs:
"DLX2: 1746
DLX5: 1749
NR2F2: 7026
SOX6: 55553
TOP2A: 7153
ZFHX3: 463"
Figure legend: "C. UMAPs depicting the relative expression of migration genes CXCR4 and EPHA5. Visualization of the gene expression of migration genes CXCR4 and EPHA5 along the pseudotime axis. D. UMAPs depicting their relative expression of neurotransmitter genes GRIA4 and GRIA2 along with visualization of their gene expression along the pseudotime axis."
NCBI gene IDs:
"CXCR4: 7852
EPHA5: 2044
GRIA2: 2891
GRIA4: 2893"
Figure legend: "B, Representative contour maps of flow cytometry for FUCCI following the loss of NF2, PTPN14, TAOK1, CREBBP, and TP53."
NCBI gene IDs:
"CREBBP: 1387
NF2: 4771
PTPN14: 5784
TAOK1: 57551
TP53: 7157"
Figure legend: "B. Vero E6 cells were transfected with 10 nM of siRNA before infection by SARS-CoV-2 at a multiplicity of infection (MOI) of 0.1 after 24 h. The numbers of viral RNA copies were quantified with RT-qPCR. The negative control siRNA served as the control and is abbreviated as &quot;Ctrl.&quot; C1-C11 represent the final candidate sequences after selection. The siRNAs capable of inhibiting up to 99% of viral envelope gene expression with P values &lt; 0.005 compared with Ctrl siRNA are marked with *. P value by Student t test."
NCBI gene IDs:
"envelope: 43740570
C1: 43740578
C11: 43740578"
Figure legend: "E. IC50 of C6 and the fully modified C6G25S. Vero E6 cells were transfected with 10, 2, 0.4, 0.08, or 0.016 nM of C6 or C6G25S before virus infection at an MOI of 0.1. The viral RNA was quantified by RT-qPCR at 24 h after infection. F. IC50 data for viral RdRp inhibition by C6G25S. Vero E6 cells were transfected with 10, 2, 0.4, 0.08, or 0.016 nM of C6G25S before virus infection at an MOI of 0.1. The viral RNA was quantified by RT-qPCR at 24 h after infection. D"
NCBI gene IDs:
"RdRp: 43740578
C6: 43740578
C6G25S: 43740578"
Figure legend: "B. IC50 for C6G25S against different variants. Vero E6 cells were transfected with 10, 2, 0.4, 0.08, or 0.016 nM of C6G25S before infection with different strains of SARS-CoV-2. The viral RNA was quantified by RT-qPCR at 24 h after infection. Data are presented as mean ± SD of three biological replicates."
NCBI gene IDs:
"C6G25S: 43740578"
Figure legend: "A, B, C. K18-hACE2-transgenic mice treated with C6G25S by 1.48 mg/L of AI for 30 min (A), 50 μL of saline containing 50 μg of C6G25S by IN (B), or PBS (C) (n = 5 per group). C6G25S distribution in lungs was visualized by in situ hybridization (ISH) staining with C6G25S-specific probe (red color). Bronchi (i) and bronchioles (ii) marked with the boxes are enlarged on the right."
NCBI gene IDs:
"C6G25S: 43740578"
Figure legend: "E, F. C57/B6 mice (n = 3 per group) were treated by AI with C6G25S 1.48 mg/L for 30 min (E) or 50 μL of saline containing 50 μg of C6G25S by IN administration (F). The C6G25S deposited in whole lungs and nasal cavities was quantified after whole tissue homogenization followed by stem-loop RT-qPCR. Quantification data represent mean ± SD. P value by Student t test."
NCBI gene IDs:
"C6G25S: 43740578"
Figure legend: "A. K18-hACE2-transgenic mice (Winkler et al., 2020) were treated once daily for 3 days before intranasal challenge with 104 plaque-forming units (PFU) of the original virus. Prophylactic treatment consists of 30 min of AI (1.48 mg/l of C6G25S), followed by IN of 50 μg C6G25S. Mice receiving vehicle control (saline) for both AI and IN are annotated as control. Viral RNA (left) and infectious virions (right) in lungs were quantified with RT-qPCR and plaque forming assay, respectively, at 2 days post-infection (dpi). B. Mice were challenged intranasally with 104 PFU of virus and co-treatment with 1.48 mg/L of C6G25S or vehicle control (saline) by AI for 30 min on day 0 (right after infection) and day 1. Viral RNA and infectious virions were quantified at 2 dpi. C"
NCBI gene IDs:
"C6G25S: 43740578"