Fig. 2

RBAT1 suppression inhibits tumor progression in vitro and in vivo. a RBAT1 was silenced in Rb and BCa cell lines by two independent GapmeRs (GapmeR1 and GapmeR2). Ctrl: negative control GapmeR. b A CCK8 assay was performed to assess cell proliferation in the control (Ctrl) and RBAT1 knockdown (GapmeR1 and GapmeR2) tumor cell lines. The results are shown as the mean ± SD in three independent experiments. *p < 0.05 and **p < 0.01. c and d A soft agar colony assay was performed to assess the tumor formation ability of RBAT1-silenced tumor cell lines (Y79, WERI-Rb-1 and 5637). Colony numbers were determined from three independent soft agar plates, the results are shown as the mean ± SD, *p < 0.05 and **p < 0.01. e Cell cycle analysis by flow cytometry was performed to determine the percentage of cells in different cell cycle phases. The percentage of cells in G0/G1 phase increased after RBAT1 knockdown in Y79, WERI-Rb-1 and 5637. X-axis represents FL2 channel-captured PI staining signals, and Y-axis represents cell counts. All histograms show the percentage (%) of cell populations from three independent experiments, the results are shown as mean ± SD. f Schematic of the orthotopic xenograft experiment. WERI-Rb-1 were injected into subretinal spaces of nude mice, which were then subjected to intravenous treatment with an RBAT1-targeting GapmeR (GapmeR1 and GapmeR2) or scrambled GapmeR (Ctrl) twice per week. g Representative image of tumor bearing eyeballs removed from the mice at the 30th day. h Top: Weights of tumors treated with GapmeR1 (n = 6), GapmeR2 (n = 6) and Ctrl (n = 6). Bottom: H&E staining to evaluate tumor formation. **p < 0.01. i Ki-67 staining of WERI-Rb-1 tumor tissues after intravenous treatment with GapmeR1, GapmeR2 and Ctrl. j Survival analysis of mice with orthotopic xenografts (WERI-Rb-1) treated with intravenous injections of GapmeR1, GapmeR2 and Ctrl twice per week; n = 6