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. 2019 Jan 11;12:168–181. doi: 10.1016/j.isci.2019.01.012

Figure 5.

Figure 5

KDM5A Is a Reprogramming Barrier

(A and B) (A) Suppression of KDM5A activity by siRNA or JIB-04 (5 μM, 48 h) promotes the enrichment of H3K4Me3 at the promoter of OCT4 in resistant fibroblasts, (B) but not that of H3K27Me3. Ct values were obtained from qRT-PCR.

(C) Suppression of KDM5A activity by JIB-04 or KDM5A siRNA (siKA) induces POU5F1, SOX2, and CDH1 mRNA levels during reprogramming, as determined by qRT-PCR.

(D) Correlation between reprogramming efficiency and KDM5A expression in HF1-HF11. The same amount of each cDNA was used as template for qRT-PCR where the Ct values indicate the expression of KDM5A.

(E) Knockdown effects of increasing concentrations of anti-KDM5A siRNA (siKDM5A, 48 h) on H3K4Me3 expression levels in HF.

(F) Reprogramming efficiency using various patient primary fibroblasts, as determined by the number of TRA-1-60-positive colonies. Cells were either transiently transfected with the indicated siRNA oligos in the first 5 days or treated with JIB-04 (5 μM, 48 h). siK5ABCD, anti-KDM5A/B/C/D siRNA; siK5AD, anti-KDM5A/D siRNA; siK5CD, anti-KDM5C/D siRNA; siK5A, anti-KDM5A siRNA; siK5B, anti-KDM5B siRNA; siK5C, anti-KDM5C siRNA; and siK5D, anti-KDM5D siRNA. In (A), (B), and (F) statistical significance was compared with OSKM-treated fibroblasts, whereas in (C) it was compared with DMSO (0.1%) treatment using two-way ANOVA and a post-hoc Tukey test. Data are represented as mean ± SD. ***p<0.001, **p<0.01, *p<0.05.