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. 2015 Feb 20;25(3):351–369. doi: 10.1038/cr.2015.22

Figure 6.

Figure 6

Δ113p53/Δ133p53 promotes DNA DSB repair by upregulating the expression of Rad51, Rad52 and Lig4. (A) Co-IP analysis of the interaction between p53 or Δ133p53 with HA-RAD51 or HA-RPA2 in H1299 cells. An anti-HA antibody was used in an immunoprecipitation. Proteins from co-IP were detected with a p53 CM1 antibody (third panel) and the HA antibody (fourth panel). The 10% of input from each sample was used as a control: top panel p53 (CM1); second panel: HA. (B) Relative mRNA expression of the listed genes in zebrafish p53M214K mutant embryos overexpressing Δ113p53, p53 or both p53 and Δ113p53 measured by qRT-PCR at 8 hpf. Gene expression was normalized against 18S rRNA and expressed as the fold change compared to the injection control. (C) Western blot analysis of proteins in human QSG7701 cells with different treatments as indicated. (D) Relative mRNA expression of the listed genes in zebrafish p53M214K mutant embryos overexpressing Δ113p53, Δ113p53R143H or Δ113p53R250W measured by qRT-PCR. (E) Effects of Δ113p53, Δ113p53R143H and Δ113p53R250W on HR, NHEJ and SSA repair frequencies. The average repair frequencies were measured by qPCR analysis of different repaired assay constructs from three repeat experiments at 10 hpf. (F) The activity of rad51, lig4 and rad52 was required for zebrafish Δ113p53-meadited HR, NHEJ and SSA repairs. The rad51-MO, lig4-MO or rad52-MO was used to knock down its corresponding gene expression in the HR, NHEJ or SSA analysis. The average repair frequencies were measured with a qPCR analysis of the repaired assay constructs from three repeat experiments at 10 hpf. Different lanes are numbered; v: versus, t-test between two lanes.