Fig. 3

Ectopic expression of Rab43 A158T in Huh7 cells enhances cell growth, colony formation and migration (a) Expression levels of endogenous Rab43 in a variety of human cancer cell lines. Western blotting confirmed an extremely low level of endogenous Rab43 expression in Huh7 cells. b Ectopic expression of wild type Rab43 and Rab43 A158T mutant in Huh7 cells. Note: the expression level of wild type Rab43 is higher than that of Rab43 A158T mutant. c Huh7 cell growth was stimulated by over-expression of Rab43 A158T mutant. Cell growth was measured by MTT assay, the growth difference was significant between empty vector and Rab43 A158T mutant transformed cells on day 5 (* p < 0.05 by ANOVA test). d-e Huh7 proliferation and colony formation was increased by over-expression of Rab43 A158T mutant. Three representative colony formation plates stained with crystal blue are presented. Cells were stained on day 10 after plating, colonies with more than fifty cells per colony were counted and the average colony number from each cell line was plotted in (e). Data are from three independent experiments and presented as average ± SD (standard deviation), *** p < 0.001 indicated statistically significant by ANOVA test. f-g Huh7 cell migration was enhanced by over-expression of Rab43 A158T mutant. Cell migration was determined by wound healing assay. The percentages of wound closure in each cell line were measured at indicated time points post wound creation. Representative wound healing images were shown in F and average wound closure rates of each cell line at the indicated times were plotted in (g). Data are from three independent experiments and presented as average ± SD (standard deviation), **p < 0.01 and *** p < 0.001 indicated statistically significant by ANOVA test