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. 2021 Apr 29;12:637825. doi: 10.3389/fphar.2021.637825

FIGURE 4.

FIGURE 4

C791-0064 inhibited RAD52 self-association. (A) Purification of human RAD52 protein analyzed by SDS-PAGE. Lane 2, total cell lysate; lane 3, supernatant; lane 4, Ni sepharose flow-through; lane 5, Ni sepharose eluate; lane 6, heparin flow-through; lane 7, heparin wash; and lane 8, heparin eluate. (B) Western blot analysis of RAD52 expressing plasmid-transformed E. coli cell lysate and purified RAD52 protein with anti-Histag antibody. (C) Schematic illustration of the electrophoretic mobility shift assay (EMSA). (D) Western blot of the EMSA result detected with anti-Histag antibody. (E) Quantification of the relative amount of polymer protein; the values were normalized by setting the value of no C791-0064, glutaraldehyde, and ssDNA reaction to be 100% (n = 3, error bars stands for SD). (F,G) Single-strand annealing efficiency of RAD52 in the presence of C791-0064. ds, double strand; ss, single strand. (H,I) Microscale thermophoresis (MST) analysis of C791-0064 (H) and G672-0031 (I) binding to Rad52 protein.