Fig. 5

TRIP13 regulates MAPK/ERK pathway via YWHAE. a RNA-seq analysis of the differentially expressed transcripts in EV and TRIP13-OE ARP-1 cells (red and blue represent high and low mRNA expression levels, respectively). Case represents ARP-1 cells stably transfected with lentivirus-mediated human TRIP13-cDNA (TRIP13-OE); Wt represents ARP-1 cells stably transfected with the empty vector (EV). GSEA indicated a positive correlation between TRIP13 overexpression and MAPK signaling activation. b Endogenous co-immunoprecipitation was conducted with H929 cells using anti-YWHAE, followed by immunoblotting using anti-TRIP13. Anti-IgG was used as a non-specific control. c and d An Analysis of TRIP13 and YWHAE expression in publicly available MM patient data sets. Increased TRIP13 or YWHAE expression is observed in plasma cells from patients with MGUS, SMM, MM and relapsed MM than from normal healthy donors. e Kaplan–Meier analyses of OS about patients from TT2 (p < 0.001) and TT3 (p < 0.05) cohorts revealed inferior outcomes among the patients with high (quartiles 4) TRIP13 or YWHAE expression compared with the remaining patients with low (quartiles 1–3) TRIP13 or YWHAE expression. f Immunohistochemical analysis of TRIP13 and YWHAE expression (positive cells are brown) in 3 representative BM specimens derived from normal and MM patients. g Immunoblotting examined that protein levels of TRIP13 and YWHAE in cells from MM patients compared with that of the normal group. h Co-immunoprecipitation of TRIP13 and YWHAE in H929 cells with or without DCZ5417. i Activity of DCZ5417 against Vector or YWHAE-OE cells were examined by CCK-8 assay. All results are expressed as means SD of three independent experiments. *P < 0.05, **P < 0.01 and ***P < 0.001 versus the control group