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. 2016 Dec 1;143(23):4368–4380. doi: 10.1242/dev.138982

Fig. 6.

Fig. 6.

Multilineage differentiation of isogenic primed versus LIF-3i-reverted hiPSC lines. (A) Differential expression of lineage-primed genes in the Polycomb (PRC2) circuit (ANOVA, P<0.001; Table S1) in seven hiPSC lines before (−) and after (+) LIF-3i reversion. Shown are heatmaps and associated log2 mean subtracted expression of PRC2 module genes of the LIF-3i-reverted versus isogenic-primed hiPSCs used in the differentiation studies below. Red, 4F-E sa-MP-iPSCs (n=4): circle, E5C3; square, E5C1; and triangle, E17C6 (or LZ6+10 for neural differentiations). Green, fibro-iPSCs (n=3): circle, C1.2; square, C2; and triangle, 7ta. *P<0.05 (paired t-tests). (B) Definitive endoderm differentiations (FOXA2+, CXCR4+ SOX17+) of isogenic LIF-3i-reverted versus primed hPSCs. Neural differentiations. (C,D) Kinetics of SOX1+ nestin+ and PAX6+ nestin+ neural progenitors in the same primed versus LIF-3i-reverted isogenic sa-MP-PSC (n=3) and fibro-hiPSC (n=3) lines described above. *P<0.05, **P<0.01 (paired t-tests). (E) Confocal microscopy of CDr3+ dye-binding neural progenitor rosettes (Yun et al., 2012). Neural rosettes were evaluated following passage of day 7 neural-induced LIF-3i-reverted (+) versus isogenic primed (−) E5C1 hiPSCs. Scale bars: 100 µm. (F) Isogenic vascular-endothelial hEB differentiations. Flow cytometry kinetics of CD31+ CD146+ (left) and KDR+ CD73+ (right) VP populations of the same isogenic sa-MP-iPSC lines as above (n=3). Error bars indicate s.e.m.