Figure 3

Depletion of XIAP increases the number of Filopodia (a) HMEC, NCI-H226 and HMEC-T cells were depleted of XIAP and immunofluorescence staining was performed using Phalloidin AlexFluor 488 to look for differences in actin structures. (b) Quantification of the number of filopodia in the three cell lines of (a) upon depletion of XIAP. Data shown are from three independent experiments with a total of 40–60 cells counted per condition (see Materials and Methods for details), P<0.001 (c) HMEC-T cells were depleted of XIAP as well as Cdc42, the quantification of the number of filopodia is shown, n=15 cells. (d) HMEC-T cells were treated with EGF stimulation after Control or XIAP siRNA treatment and immunofluorescence was performed using three different stains: Phalloidin for Actin, Hoechst for the nucleus and a Cdc42 primary antibody followed by Cy3 mouse secondary to check for localization of Cdc42 protein in the cell (e) Quantification of the number of filopodia seen in Figure (d) with n=10 cells counted per condition, P<0.01