Figure 6.

Zyxin silencing inhibited VSMC actin cytoskeleton reorganization. (A) silencing of zyxin inhibited VSMC spreading. Non-silencing control or zyxin-shRNA transfected VSMCs were suspended with serum-free culture medium and plated on 50 μg/ml fibronectin-coated dishes for 1, 3, and 5 h under standard culture conditions. Cells were then fixed and stained with Alexa 568 conjugated-Phalloidin. Images were taken with a 40x oil objective. Scale Bars, 30 μm. (B) Bar graphs represent mean of cell spreading areas calculated using NIH Image J (^*p < 0.05 vs. control). Error bars represent error of the mean. Results are representative of five independent experiments. (C) Silencing of zyxin inhibited jasplakinolide-induced actin-filament strengthening. (D) Bar graphs represent mean of actin-staining fluorescence intensity of individual cells. CK4-DMSO, n = 15; CK4-JSP, n = 13; zyxin-DMSO, n = 13; and zyxin-JSP, n = 13. VSMCs stably transfected with non-silenced control or zyxin-shRNA were plated with regular culture medium on 50 μg/ml fibronectin-coated dishes overnight and cells were treated with 100 nM Jasplakinolide for 1 h under standard culture conditions. Cells were then fixed and stained with phalloidin. The fluorescence intensity of individual cells was calculated as the sum of a z-stack trans-focus images that were captured using confocal microscopy. Scale bars, 20 μm. (^*p < 0.05 vs. control). Results are representative of three independent experiments.