Fig. 4: Stage-specific regulation of TFAP2C during EVT differentiation. | Nature Communications

Fig. 4: Stage-specific regulation of TFAP2C during EVT differentiation.

From: The transcriptional regulatory network modulating human trophoblast stem cells to extravillous trophoblast differentiation

Fig. 4

a Experimental design for time-course TFAP2C KD (top) and OE (bottom). b Relative expression of TFAP2C and EVT marker genes in EVTs after KD of TFAP2C at different time points. E: KD day −1–8, M: KD day 3–8, L: KD day 5–8. Error bars: mean ± SD (n = 3, independent repeats). Significance by two-sided Student’s t-test. c Invasion ability of EVTs after KD of TFAP2C at different time points and individual control cells. Early: KD day −1–8, Mid: KD day 3–8, Late: KD day 5–8. Scale bar: 100 µm. d Immunofluorescence of EVT marker genes in EVTs after TFAP2C KD at different time points. Scale bar: 100 µm. Two independent repeats were carried out, resulting in similar results. e Brightfield images of EVTs overexpressing TFAP2C (Dox-treated day 2–8) and control. Scale bars: 100 µm. Two independent repeats were carried out, resulting in similar results. f Relative expression of EVT markers in TFAP2C OE compared to each control. Dox was treated on day 2, and cells were collected on days 3, 5, and 8. Error bars: mean ± SD (n = 3, independent repeats). Significance by two-sided Student’s t-test. g Immunofluorescence of HLA-G and TP63 in TFAP2C OE (day 2–8) and control. Scale bar: 100 µm. Two independent repeats were carried out, resulting in similar results. h Box plot illustrates class 1–4 gene expression in TFAP2C OE (day 2–8) compared to control (***p < 2.2e-16, two-tailed Wilcoxon rank-sum test). Each group has two independent RNA-seq results. The median (black bar) and box (25th to 75th percentiles) represent data distribution, while whiskers denote the minimum to maximum values. i, GSEA of TFAP2C OE and control using the EVT and CT marker gene sets defined in scRNA-seq data of human first-trimester placenta19,102. NES normalized enrichment score, FDR false discovery rate. j Correlation heatmap showing comparisons of TFAP2C OE with EVTs after KD of individual EVT regulators. Spearman correlations were calculated using log2-fold changes of the class 1–4 genes.

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