Fig. 5: DNA damage response of primed vs. naïve DVP. | Nature Communications

Fig. 5: DNA damage response of primed vs. naïve DVP.

From: Vascular progenitors generated from tankyrase inhibitor-regulated naïve diabetic human iPSC potentiate efficient revascularization of ischemic retina

Fig. 5

a purified and expanded DVP or N-DVP (E1CA2) were treated with the radiomimetic drug NCS for 5 h before fixation and staining with antibodies for detection of human CD31+ cells and phosphorylated H2AX (pH2AX) positive nuclear foci (i.e., DAPI co-staining) to reveal double-strand DNA breaks (arrows); Scale Bar = 50 μm. b Quantification of pH2AX foci per nuclei in isogenic DVP vs. N-DVP with or without induction of NCS DNA damage. Shown are numbers of DAPI+ nuclei per field with no pH2AX foci (green), and DAPI+ nuclei with 1–5 foci (light pink), 6–10 (dark pink) and >10 pH2AX foci (red). ***p < 0.0001; Chi-Square tests (c) Lysates of primed DVP and N-DVP (E1C1) cultured in EGM2 and treated with (+) or without (−) NCS were analyzed by Western blotting for expressions of proteins activated by DNA damage and apoptosis (i.e., total H2AX and phosphorylated H2AX (P-H2AX), RAD51, RAD54, phosphorylated p53 (P-p53), total DNA-PK, and phosphorylated DNA-PK (P-DNA-PK).

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