FIGURE 5.
ELMO2 knockdown impairs microtubule capture at the membrane during integrin-mediated cell spreading. A–D, MDA-MB-231 cells transfected with Myc alone (A), HA-ACF7 (B), Myc-ELMO1 and HA-ACF7 (C), and Myc-ELMO1PxxP and HA-ACF7 plasmids (D) together with either a siRNA targeting ELMO2 or a nonspecific siRNA were serum-starved. The cells were plated on fibronectin-coated chambers for 2 h and stained for β-tubulin, Myc, HA, and DAPI. The main panels show the β-tubulin staining, whereas the insets are the overlays of the staining for ELMO1, ACF7, and DAPI. Scale bar, 20 μm. E, quantification of the percentage of cells with microtubule reaching the cellular cortex in the various conditions (**, p < 0.005; by ANOVA test and Bonferroni's multiple comparison; error bars represent S.E., n = 3).