Figure 7.

RNF8 can inhibit p53‐dependent apoptosis by attenuating Tip60‐dependent excessive activation of ATM. A, (i). U2OS cells were transfected with indicated plasmids for 48 h, then treated with or without Eto (10 μmol/L). The expression of the indicated proteins was showed by immunoblot. β‐Actin was used as the loading control. (ii) The quantification of p53‐120ac, p53 and γ‐H2AX protein levels. **P < .01, n ≥ 3. B, U2OS cells were transfected with indicated plasmids for 48 h, and total RNA was extracted and reversely transcribed into cDNA, and the expression of related genes was detected by PCR. C, (i) U2OS cells were transfected with indicated plasmids for 48 h and then incubated with 0, 50 μmol/L of Eto for 24 h, and the flow cytometry analysis of Annexin V‐FITC/PI staining was conducted to examine the dead cells. (ii) The total number of Annexin V‐positive and Annexin V/PI double‐positive cells was quantified. *P < .05, n ≥ 3. D, Different types of HEK‐293T cells were transfected with indicated plasmids for 48 h, and total RNA was extracted and reversely transcribed into cDNA, and the expression of related genes was detected by PCR. E and F, Different types of HEK‐293T cells were transfected with indicated plasmids for 48 h; then, the cells were treated with Eto (10 μmol/L) for 20 min and repaired 1 h. The immunoblot showed the expression of the indicated proteins. β‐Actin and GAPDH were used as the loading control