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. 2021 Mar 25;11:646256. doi: 10.3389/fonc.2021.646256

Figure 4.

Figure 4

RFWD3-dependent Rad51 ubiquitination is involved in the VPA-mediated bidirectional effect. (A) RFWD3 protein expression was detected with VPA and IR treatment. (B) The interaction between RFWD3 and Rad51 proteins was detected by protein beads containing Rad51 antibody through Western blot. (C) The foci formation of RFWD3 and Rad51 in this paired cell was detected at 6 h post-8 Gy IR. Co-localization of RFWD3 and Rad51 proteins was shown in the figure. The scale bar represented as 10µm. (D) siRNA was designed to abolish the function of RFWD3 and the expression of Rad51 under those treatments was detected by Western blot. (E) Ubiquitination of Rad51 on transformed and normal MCF10A cells after RFWD3 depletion was detected by Western blot. Bands in the graphs were quantified by ImageJ software (Wayne Rasband). Each data point in the graph was from three independent experiments (mean ± SD); P-values were calculated by t-test (**P < 0.01).