Figure 5.

AA Establishes a Requirement for EHMT Activity during Enhanced iPSC Reprogramming

(A) Alkaline phosphatase staining of transgene-independent iPSC colonies obtained after reprogramming MEFs in the presence of the indicated compounds in the absence or presence of UNC0638.

(B) Quantification of iPSC colonies formed under the indicated conditions. n = 3 independent reprogramming experiments.

(C) Quantification of iPSC colonies formed in the presence of the indicated dual-compound conditions.

n = 3 independent reprogramming experiments. Significance in (B and C) was calculated by two-way ANOVA with Sidak post test with adjusted p values of ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001, and ^∗∗∗∗p < 0.0001.

(D) Schematic of the approach taken to test for potential functional deficits of iPSCs caused by transient EHMT inhibition during reprogramming.

(E) Quantification of viable pups obtained after blastocyst injection with the indicated iPSCs. Each circle represents the offspring obtained from one recipient female (n = 10 for each condition) of 20 injected blastocysts. ^∗∗p < 0.01 with Mann-Whitney test. Lines show mean with SD.

(F) Heatmap of Pearson's correlation coefficient for all expressed genes between three iPSC cultures derived in the presence of UNC0638 under 3c conditions and three iPSC cultures derived in the absence of UNC0638.

(G) Heatmap of the expression levels (RPKM) of the 328 ISGs (see Figure 3C) in established iPSCs derived in either the absence or the presence of UNC0638.