Fig. 2: Knockdown of UBAC2 represses the proliferation of BC cells in vitro and in vivo.

a, b The efficiency of UBAC2 knockdown in EJ and UMUC3 cells was detected by qRT-PCR (a) and western blotting assay (b). β-actin was used as loading control. c Flow cytometry indicated that knockdown of UBAC2 resulted in cell cycle arrest at G0/G1 phase in both EJ and UMUC3 cells compared with negative control. d Cell viability was evaluated by Cell Counting Kit-8 assay. e Plate colony formation assay showed that the colony formation activity was inhibited after shUBAC2 transfection. Colonies with more than 50 cells were counted. f Tumors collected from mice were exhibited after 1 month of hypodermic injection. g EJ cells stably transfected with shUBAC2 plasmids or control plasmids were subcutaneously injected into the right flanks of the nude mice (2 × 10⁶ cells per mouse, n = 5 for each group). h The volumes of tumors were measured at 1–4 weeks after hypodermic injection and the growth rates of xenograft tumors upon shUBAC2 or shNC treatment were analyzed. i The weights of xenograft tumors were measured and analyzed after one month of hypodermic injection. j Ki67 staining were performed to show the proliferation index in xenograft tumors. The images were analyzed by calculating the integrated optical density per stained area (IOD/area). Data are presented as means ± SEM from three independent replicates or as means ± SEM of five mice from each group. **p < 0.01, ***p < 0.001 (Student’s t test).