Fig. 1: Gm11549 is a direct target gene of Nodal signaling in mesendoderm differentiation of mESCs. | Nature Communications

Fig. 1: Gm11549 is a direct target gene of Nodal signaling in mesendoderm differentiation of mESCs.

From: A Nodal enhanced micropeptide NEMEP regulates glucose uptake during mesendoderm differentiation of embryonic stem cells

Fig. 1: Gm11549 is a direct target gene of Nodal signaling in mesendoderm differentiation of mESCs.The alternative text for this image may have been generated using AI.

a Volcano plot showing differentially accumulated long noncoding RNAs from transcriptome datasets of day 2.5 murine EBs treated with either Activin A or SB431542 for 2 h (n = 2 biologically independent samples). Blue, long noncoding transcripts downregulated in Activin A -versus SB431542-treated EBs (fold change < 0.5, p < 0.05); Red, long noncoding transcripts upregulated in Activin A- versus SB431542- treated EBs (fold change > 2, p < 0.05) (GEO: GSE115169). b Gene track view of RNA-seq for Activin A (AC)- and SB431532 (SB)—treated day 2.5 EBs for 2 h; SMAD2/3 ChIP-seq, SMAD4 ChIP-seq, TRIM33 ChIP-seq, and FOXH1 ChIP-seq in AC- and SB- treated day 2.5 EBs for 2 h (GSE70486, GSE125116); FANTOM5 CAGE at the Gm11549 locus. Three forms of Gm11549 transcript (Genbank, Refseq, and our RACE from days 3 EBs) are shown. c qPCR analysis of Gm11549, Lefty1, and Lefty2 expression in Activin A (AC)—or SB431542 (SB) - treated day 3 EBs for 2 h with or without pretreatment with cycloheximide (CHX) for 1 h. d qPCR analysis of the indicated transcripts in day 0 to day 6 EBs of in vitro differentiation. Fgf5 is a primary ectoderm marker. Mixl1, Gsc, and T are markers for the primitive streak. Foxa2 is a marker for the definitive endoderm. e Adapted from47: Corn plots showing the spatial pattern of Gm11549 expression in E7.0 mouse embryos. A: anterior; P: posterior; L: left lateral; R: right lateral. f qPCR analysis of Gm11549 expression in day 0 to day 5 EBs from WT or Smad2 KO, Smad3 KD, Smad2 KO/Smad3 KD, and Smad4 KO cells. g qPCR analysis of Gm11549 expression in day 0 to day 6 EBs from WT or Trim33 KO cells. h qPCR analysis of Gm11549 expression in day 0 to day EBs from WT or Foxh1 KD cells. i qPCR analysis of Gm11549 expression in day 0 to day 5 EBs from in WT, Cripto KD, Cryptic KD, Cripto KD/Cryptic KD cells. c, d, f–i: Data are the mean ± S.E.M, n = 4 biologically independent samples. P values were determined by one-way (d) with Dunnett’s corrections or two-way ANOVA with Tukey’s corrections (c, f, i) or with Sidak corrections (g, h), and data are representative of three independent experiments with similar results. Source data are provided as a Source Data file.

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