Figure 4.

NMP-derived cells stabilize as epithelial PNPs and NC progenitors

(A and B) Normalized expression levels of known markers of NMPs, PNPs, and NPs at 36 h (A) and in P5 epithelial- and mesenchymal-enriched samples (B) as determined by RNA-seq. Error bars show SEM (n = 3 independent differentiations). ^∗∗∗FDR < 1%, a FC of at least ±2 compared with epithelial-enriched samples, and a base mean > 100.

(C) Representative immunostaining of Brachyury (green), SOX2 (red), and PAX6 (magenta) confirming the expression patterns shown in (A) and (B). Scale bars, 100 μm.

(D) Volcano plot showing differential expression between epithelial (blue) and mesenchymal (red) cells at P5.

(E and F) Representative immunostaining of NC markers SNAI1, SOX1 and SOX9 (E), and ETS1 (F) co-stained with epithelial PNP marker CDX2 (gray) and DAPI (blue). Scale bars, 100 μm.

(G) Log₂ FC (versus hESCs) of NC marker genes in P5 mesenchymal- and epithelial-enriched samples compared with previously published trunk NC microarray data (Frith et al., 2018).