Fig. 3
From: Cell shape information is transduced through tension-independent mechanisms
Integrin β3 controls shape-driven phenotype in podocytes. a Representative images of podocytes plated on unpatterned and micropatterned surfaces, and stained either for (left) nephrin (green) and F-actin (red), or (right) podocin (green) and F-actin (red). Both podocin and nephrin were highly localized within peripheral processes. Treating cells with integrin β3 blocking antibodies abolished the localization effect. Podocytes treated with integrin β1 blocking antibodies exhibit limited spreading; however, podocin and nephrin phenotype was relatively unaffected (insets show the autofluorescent patterns for clarity). b Quantitative analysis of whole-cell nephrin and podocin intensities in patterned and unpatterned podocytes treated with either β1 or β3 blocking antibodies. Values given as mean ± SEM; n = 80 chosen randomly from eight different slides cultured independently (*p < 0.01 vs. UNP; one-way ANOVA followed by a post hoc Tukey test). c No significant differences in integrin expression were observed between fibronectin coated and uncoated surfaces; spatial integrin expression did not depend on the ECM coating in 3-D micropatterned podocytes. Integrin β1, α5, β3, and αv (cyan, stained independently), fibronectin (green), and F-actin (red) in podocytes plated on channel surfaces with and without fibronectin coating. d Podocytes were treated with varying concentrations of blebbistatin for 12 h prior to fixation and stained for (top) nephrin (green) and F-actin (red), (middle) p-FAK (cyan) and F-actin (red), or (bottom) p-myosin (green) and F-actin (red). Phospho-myosin intensity decreased gradually with increasing blebbistatin concentration, whereas recruitment of p-FAK to focal adhesions was not affected by blebbistatin up to 10 μM
