Fig. 1

Biphasic roles of RhoGDIβ in UROtsa cells after different BBN exposure lengths. A Treatment of UROtsa cells with 400 μM BBN for times indicated. The cell lysates prepared at each time point were evaluated RhoGDIβ expression using Western blot. The bands of RhoGDIβ and β-actin were not derived from the same membrane due to big differential binding intensities of two antibodies. B, C UROtsa^C2mo and UROtsa^BBN2mo cells were tested in a soft agar assay in the presence or absence of EGF (20 ng/ml). Representative images of colonies of indicated cells are shown; results are presented as colonies/10⁴ cells seeded. Bars represent mean ± SD of three independent experiments. The asterisk indicates significant difference vs. control group (p < 0.05). D, E UROtsa^C6mo and UROtsa^BBN6mo cell invasion (Invasion Chamber). The asterisk indicates significant difference between cells. F Ectopic RhoGDIβ constructs were stably transfected into UROtsa^BBN2mo cells, and the over-expressed efficiency of RhoGDIβ protein was assessed by using Western blot. G, H Cell anchorage-independent growth of UROtsa^BBN2mo/Vector and UROtsa^BBN2mo/GFP-RhoGDIβ cells was determined in soft agar in the presence of EGF (20 ng/ml). Representative images of colonies are shown. The asterisk indicates significant difference vs. vector transfectants (p < 0.05). I RhoGDIβ knockdown constructs were stably transfected into UROtsa^BBN6mo cells, and knockdown efficiency was then determined using Western blot. J, K UROtsa^BBN6mo/Nonsense and UROtsa^BBN6mo/shRhoGDIβ#2 transfectants were subjected to transwell invasion assay. The asterisk indicates significant difference in comparison to UROtsa^BBN6mo/Nonsense cells