Figure 2
Mena11a expression maintains junctional integrity.
(A–E): MCF7 cells. (A) Immunofluorescence showing endogenous ZO-1 and Mena11a localization. Scale bar, 10 μm. (B) Immunofluorescence showing endogenous E-cadherin and Mena11a localization. Scale bar, 10 μm. (C) Western blot analysis. Membranes probed with anti Mena11a and anti pan-Mena antibodies. α-Tubulin: loading control. (D) Quantitative analysis of relative ratio of Mena11a:α-Tubulin, determined by densitometry. Fold change in expression is relative to appropriate control. Error bars: SEM. Results represent triplicates. (E) (Left panel) 3D-SIM images showing E-cadherin localization in MCF7 cells with isoform-specific knockdown of Mena11a, using two different shRNAs (sh-1, sh-2) and control shRNAs (sh-1C). Space-filling GFP in blue indicates cells expressing Mena11a shRNAs or control shRNAs. F-actin is visualized by phalloidin labeling. Insets: 7X magnification. Scale bar, 10 μm. (Right panel, top) Quantitation of junctional E-cadherin fluorescence intensity. a.u. = arbitrary units. >30 cells analyzed. Error bars: SEM. Results represent triplicates. One-way ANOVA *p < 0.05, n.s., not significant. (Right panel, bottom) Lateral distribution of junctional E-cadherin. >30 cells analyzed. Error bars: SEM. Results represent triplicates. One-way ANOVA n.s., not significant.
