Fig. 3

DCZ5417 specifically binds and inhibits TRIP13 in Cells. a The structure of DCZ5417 (green) and its binding mode to TRIP13 (gray) as determined by molecular docking. b The binding mode of DCZ5417, DCZ5419 and NCTD with TRIP13 determined by molecular docking. c SPR biosensor was used to detect the binding of DCZ5417 to TRIP13. d A pull-down assay was used to detect the binding of DCZ5417-biotin to TRIP13. Anti-MM activity of DCZ5417 and DCZ5417-biotin was compared (below). e Cellular thermal shift assay to examine interactions of DCZ5417 (10 µmol/L) with TRIP13. f Relative ATPase activity was examined after DCZ5417 and NCTD treatment by ADP-Glo™ Kinase Assay. g The viability of MM cells transfected with empty vector or TRIP13-sgRNA with DCZ5417 treatment (0, 2.5, 5, 10 and 20 µmol/L, 48 h) was analyzed by a CCK-8 assay. SgControl represents nontarget scramble-transfected cells. TRIP13 sgRNA represents TRIP13-silenced cells. The result is expressed as means SD of three independent experiments. h CCK-8 assay was performed on TRIP13 OE cells or empty vector-transfected cells. Immunoblotting was used to examine protein level of TRIP13 in TRIP13 OE and Vector cells. Vector represents nontarget scramble–transfected cells. TRIP13 OE represents overexpression of TRIP13 in cells. Data are presented as the means ± SD of 3 independent experiments. *P < 0.05, **P < 0.01 and ***P < 0.001 versus the control group