Fig. 3

RhoH depletion reduces cell migration. a RhoH depletion reduces migration on uncoated plastic or Matrigel-coated plastic. PC3 cells were transfected with a control siRNA pool or the pool of four siRNAs targeting RhoH. After 24 h, cells were seeded around Oris™ stoppers. After 24 h, stoppers were removed and migration was determined 24 h later. Left, example images of cells 24 h after removal of stoppers. White dotted circle indicates wound area at 0 h. Right, graph showing migration as % of siControl +/− SEM, n = 3; **p ≤ 0.01, Student’s t test. b Individual siRNAs targeting RhoH reduce migration. PC3 cells were transfected with indicated siRNAs. Cell migration was determined as in a. Graph shows migration as % of siControl +/− SEM, n = 3; **p ≤ 0.01, ***p ≤ 0.001, Student’s t test. c Relative mRNA expression levels following RhoH knockdown, compared to control siRNA-transfected cells. Levels were measured by quantitative real-time PCR (qPCR). n = 3; **p ≤ 0.01, ***p ≤ 0.001, Student’s t test. d Rho siRNAs reduce expression of exogenous RhoH. PC3 cells were transfected with the indicated siRNAs. Cells were transfected with human astrocyte (HA)-RhoH 48 h after siRNA transfection. After 24 h, cell lysates were analysed by western blotting with an anti-RhoH antibody. α-tubulin was used as a loading control. e RhoH depletion reduces migration through transwells. PC3 cells were transfected with the indicated siRNAs. Cell migration was determined through transwells with 1% fetal calf serum (FCS) as chemoattractant, and migration index calculated relative to siControl-transfected cells +/− SEM, n = 3; ***p ≤ 0.001, Student’s t test