Figure 4. RhoA/p160ROCK pathway regulates actin stress fiber formation in PhLP3 siRNA silenced CHO LB01 cells.

A commercially available RhoA-GTP/Rhotekin pull down assay was carried out on siRNA silenced PhLP3 LB01 cells. Samples were collected 24 h, 48 h and 72 h post transfection and activated RhoA was detected at 72 h post-knockdown with an anti-Rho A antibody but not in plasmid only control cells (A). LB01 cells transfected as in panel A for 72 h were then treated for 24 h with the ROCK inhibitor Y2763 (either 3 µM or 10 µM), fixed and stained for actin (rhodamine phalloidin, red). F-actin disassembly was observed at both Y2763 concentrations and the concentration of peripheral actin was observed with 10 µM Y2763 (B).