Fig. 4: GGPP-mediated geranylgeranylation of RhoB protein is associated with the migration ability of BLCA cells.

a Flowchart for the detection of MVA pathway intermediate metabolite (FPP and GGPP) levels and proteomic changes in BLCA cells after MVA pathway inhibition by ZOL or simvastatin. The contents of FPP (b) and GGPP (c) in T24 cells treated with ZOL (20 μM) (n = 3) or simvastatin (5 μM) (n = 3) were measured by LC–MS/MS analysis. d Heatmaps of proteins whose expression significantly changed in T24 cells after MVA pathway inhibition by ZOL or simvastatin, respectively. e Western blotting was performed to detect RhoB protein in BLCA cells after treatment with ZOL. GAPDH was used as the loading control. Representative images (f) and statistical graph (g and h) of transwell migration assays from the indicated groups after treatment of BLCA cells (UM-UC-3 and T24) with RhoB siRNA and ZOL (20 μM), respectively, or in combination (n = 3). The scale bar is 200 μm. i Schematic representation of BLCA cells treated with ZOL (20 μM), GGPP (5 μM) and GGTI298 (10 μM), alone or in combination. Representative images (j) and statistical analysis (k and l) of transwell migration assays from the indicated groups after treatment of BLCA cells (UM-UC-3 and T24) with ZOL (20 μM), GGPP (5 μM) and GGTI298 (10 μM), separately or in combination (n = 3). The scale bar is 200 μm. Changes in RhoB protein expression in different groups of UM-UC-3 (k) and T24 (l) cells were also detected by western blotting. GAPDH was used as the loading control. The n number represents n biologically independent experiments in each group. Statistical significance was ascertained by two-tailed unpaired Student’s t-test (b, c, k and l) and one-way ANOVA with Dunnett’s multiple comparisons test (g and h). The data are shown as the mean ± SD.