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. Author manuscript; available in PMC: 2015 Sep 30.
Published in final edited form as: Cell Metab. 2015 Aug 4;22(2):239–252. doi: 10.1016/j.cmet.2015.07.015

Figure 7. Rescue of DNA Damage Checkpoint Proteins by miR200 Knockdown in Human Medalist +C Fibroblasts and iPSCs.

Figure 7

(A and B) Quantitative real-time qPCR analysis to assess combined knockdown (KD) of miR200A, miR200B, and miR200C in (A) iPSCs of Medalist +C patients (N = 6) or (B) Medalist +C fibroblasts (M6, M9, and M12; N = 3).

(C) Quantitation of reprogramming efficiency of human Medalist +C fibroblasts (M6, M9, M12) treated with scrambled siRNA or miR200 siRNA (N = 3).

(D) Immunoblot analysis highlighting the restoration of ATM/ATR proteins in Medalist +C fibroblasts (M6, M9, M12) treated with miR200 siRNA compared to scrambled siRNA (N = 3).

(E) Immunoblot analysis showed decreased expression of pH2AX/cleaved caspase-3 in Medalist +C fibroblasts treated with miR200 siRNA (N = 3).

(F) Immunostaining analysis of pH2AX in miR200 siRNA-treated Medalist +C human iPSCs as compared with scrambled siRNA-treated iPSCs. pH2AX, green; DAPI, blue. Scale bar: 50 µm.

(G) Quantitative analyses of immunostaining of pH2AX in human iPSCs (N = 6) derived from Medalist +C patients and subjected to siRNA-mediated knockdown of miR200 (72 hr).

(H) Quantitative real-time PCR analysis showing the expression of hTERT gene in control (cntrl1, cntrl2), Medalist −C (M4, M10), and Medalist +C fibroblasts (M6, M12) (N = 2).

All data are presented as mean ± SD. *p value < 0.001.