|
|
|
|
|
|
|
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
|
Fig. 7. Phosphorylation of LIMK1 and cofilin is not required for EGF-stimulated lamellipod protrusion and barbed end formation following stimulation with EGF. (A) MTLn3 cells were pre-incubated with the ROCK inhibitor Y-27632 (10 µM) for 30 minutes and fixed at 0 and 60 seconds following stimulation with EGF to inhibit the phosphorylation of both LIMK1 and cofilin. Panel shows representative images of the F-actin cytoskeleton stained with Alexa Fluor-488-labeled phalloidin in control and Y-27632-treated cells. (Bar, 10 µm) (B) The change in lamellipod protrusion area is unaffected by ROCK inhibition. MTLn3 cells were treated with or without the ROCK inhibitor, Y-27632 and fixed at 0 and 60 seconds after stimulation with EGF. The average fold change in cell area measured at 0 and 60 seconds EGF in control and Y-27632-treated cells is identical. Student's t-test showed no statistical difference in lamellar protrusion between control and Y-27632 cells. (Error bars indicate the s.e.m.) (C) Time lapse microscopy of MTLn3 cells demonstrate no change in the initial rate or final extent of lamellipod protrusion in Y-27632-treated cells (
) compared with control cells (
) following stimulation with EGF. (D) The change in the appearance of new barbed ends is unaffected by ROCK inhibition. Time lapse microscopy of ß-actin-GFP MTLn3 cells demonstrate there is no effect on the fold change in the appearance of new barbed ends in Y-27632-treated cells following stimulation with EGF compared with control cells. For panels C and D, error bars indicate the s.e.m. for nine control cells and 11 Y-27632-treated cells (n=2 separate experiments).
|
