Fig. 8

Salinomycin inhibited VM through inhibition of Rho-GTPases. a Morphology of SKBR3-TR cells incubated with DMSO or 4 μM salinomycin for 2 h. Representative confocal microscopy photographs of cells stained with Alexa Fluor 488-conjugated phalloidin and DAPI are shown. Scale bar, 20 μm. b Sizes of SKBR3-TR cells treated with DMSO (n = 37; mean ± SD) or 4 μM salinomycin (n = 50; mean ± SD). c Box plots of the roundness of SKBR3-TR cells treated with DMSO (n = 37) or 4 μM salinomycin (n = 50). The whiskers represent the 10th–90th percentiles. P, Mann-Whitney U test. d Box plots of the integrated density, which represents the amount of F-actin per cell, of SKBR3-TR cells treated with DMSO (n = 37) or 4 μM salinomycin (n = 50). The whiskers depict the 10th–90th percentiles. P, Mann-Whitney U test. e Cell migration assay showing the gap width at 36 h (percentage of gap width at 0 h) for SKBR3-TR cells treated with DMSO or 1 μM salinomycin (n = 3; mean ± SD). f Pulldown assay of GTP-bound Rho-GTPases extracted from JIMT-1 cells treated with DMSO, salinomycin, or salinomycin plus Rho Activator II. PD, pulldown; IB, immunoblot. g Cell migration assay using JIMT-1 cells treated with DMSO, 0.5 μM salinomycin or 0.5 μM salinomycin plus 2 μg/mL Rho Activator II (n = 3; mean ± SD). h Representative photographs showing tube formation in JIMT-1 cells treated for 24 h with DMSO, 0.3 μM salinomycin or 0.3 μM salinomycin plus 1 μg/mL Rho Activator II. Scale bar, 0.5 mm. i The numbers of tubes formed are shown (n = 3; mean ± SD)