Oxidative Medicine and Cellular Longevity / 2019 / Article / Fig 3

Research Article

Overexpression of BRCA1 in Neural Stem Cells Enhances Cell Survival and Functional Recovery after Transplantation into Experimental Ischemic Stroke

Figure 3

BRCA1 interacted with p53 and suppressed p53-associated apoptotic pathway. (a) The lysates from OGD/R-treated NSCs were immunoprecipitated with anti-BRCA1 or anti-p53. Then, total lysates and immunoprecipitates were subjected to immunoblot analysis. . (b) GST-BRCA1 fusion proteins containing different domains of BRCA1 were purified and incubated with OGD/R-treated NSC lysates. Protein to protein interaction was detected between RING finger domain and p53 as well as BRCT domain and p53. . (c) Transfected OGD/R NSCs were pretreated with 50 μM MG132 for 6 h, and then, cell lysates were immunoprecipitated with anti-p53 antibody. The level of p53 ubiquitination was analyzed by western blotting. . (d) Western blot and quantitative analysis of p53 proapoptotic pathway 24 h after reoxygenation. Abolished upregulation of p53, Bax, and cleaved caspase-3 by OGD/R induction and reversed decrement of Bcl-2 by OGD/R induction were observed in BRCA1-overexpressed NSCs. . Data are expressed as ; , , and vs. the control group; and vs. the OGD/R group.