(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.