Fig. 5.
Expression of constitutively active RhoA inhibits Fsk-induced changes in LNCaP cell morphology in a dominant-negative manner.
Panel A: LNCaP cells were plated into 6-well plates at a density of 3 × 105 cells per well and transfected with 1 μg of either pRK5, myc.WTRhoA or constitutively active myc.Q63LRhoA as described above. Cells were then stimulated with vehicle (0.1% (v/v) EtOH) and 10 μM Fsk and images captured as described at 0 h (0 h) and 1 h post-stimulation.
Panel B: Changes in LNCaP morphology consistent with NE-like differentiation were assessed via increases in mean dendrite length. Results are represented as mean values ± SEM for n = 3 separate experiments. ***p < 0.001 vs. 0 h, ###p < 0.001 vs. vehicle, +++p < 0.001 vs. pRK5.
Panel C: Expression of RhoA mutants was assessed via 9E10 immunoblotting for the myc epitope. * indicates a non-specifically labelled immunoreactive band that serves as a control for equal protein loading.