Stiff Substrates Enhance Endothelial Oxidative Stress in Response to Protein Kinase C Activation

Figure 5

PMA treatment did not increase pMLC localization to actin stress fibers in cells on varied stiffness substrates. PAEC monolayers on 6, 14, or 29 kPa gels were treated with 1 μM PMA for 15 or 30 minutes, prior to fixation and pMLC immunofluorescent labeling. For the positive control, cells on a 29 kPa gel were treated with 10 U/mL thrombin for 30 minutes. Images are maximum intensity projections from 60x confocal -stacks. Scale bar is 25 μm. pMLC-positive pixels were quantified using the custom MATLAB code. Stiffness and PMA treatment were not significant by -way ANOVA.