High-Fidelity Reprogrammed Human IPSCs Have a High Efficacy of DNA Repair and Resemble hESCs in Their MYC Transcriptional Signature

Figure 2

sa-CB-iPSCs closely resemble hESCs in DSB damage response to radiation. ((a)(i), (a)(ii), and (b)) Representative Western blot analysis depicting the expression of phosphorylated ATM (pATM) and H2AX (γH2AX) in cell lysates from H9, ES03, CB6.2, iCB9, and iHuF3 at time 0 and at 1 h, 2 h, and 4 h after IR. β-Actin and β-tubulin were used as loading controls. Cells were exposed to IR (X-ray; 2 Gy) recovered at the indicated time points and immunoblotting was performed to analyze the kinetics of DDR protein expression. ((c) and (d)) Densitometry analysis of the Western blots for (c) γH2AX and (d) pATM (normalized to total ATM), using ImageJ software. Statistical significance of the data was determined using 2-way ANOVA analysis with Bonferroni posttests to compare the replicates (three independent experiments). γH2AX expression in iHuF3 is significantly different at 4 h compared to the following (versus H9 and CB6.2, ). pATM expression in iHuF3 is significantly different at 1 h and 2 h, compared to all other cell lines ().

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