A. Immunoblot of cortactin expression (top panels) and β-actin loading control (bottom panels) in SCC61, HeLa and MCF10A stable (3 left panels) or transient (2 right panels) KD cell lines. For the transient siRNA experiments NTC indicates nontargeting control and CTTN indicates cortactin-specific siRNA. B. Representative images and analyses of SCC61 cells (top), HeLa cells (middle) and MCF10A cells (bottom) stably expressing control shRNA (scrambled), human cortactin-specific shRNA (KD1, KD2), or cortactin-KD1 plus shRNA-resistant wild-type mouse cortactin (rescue) and immunostained for the Golgi marker, GM130 (green), actin filaments (rhodamine-phalloidin, not shown) and the nucleus (Hoechst, blue). Graphs show quantification of the ratio of Golgi area to total cell area of individual cells. C. Representative images and analyses of transient control (NTC) or cortactin-KD (CTTN) SCC61 or HeLa cells. Scale bar = 25 µm. For each condition, n≥20 cells from each of 3 independent experiments. Data are presented as box and whiskers plots with the box indicating the 25th and 75th percentiles, solid line indicating the median, and the whiskers indicating the 95% confidence intervals.*p<0.05.