Fig. 2

A) Confocal images of HeLa cells stained for PI3K-C2α (green) and a-tubulin (red) (left) during cytokinesis and (right) fluorescence intensity along the line showing localization of PI3K-C2α at midbody. (B) Hela cells transfected with wild-type, kinase inactive (KD), clathrin-binding deletion (D1-380), and PX-binding mutant PI3K-C2α GFP-tagged. Immunofluorescence staining by using antibody to GFP showing (left) the enrichment of the different PI3K-C2α constructs and (right) their quantification at the midbody. (C) Confocal images of (left) HeLa cells stained for PI3K-C2α (green), 4′,6-diamidino-2-phenylindole (DAPI) (blue), and g-tubulin (red) and (right) fluorescence intensity along the line showing colocalization between PI3K-C2α and g-tubulin. (D) Immunoprecipitation of myc-PI3K-C2α from cells synchronized in cytokinesis. (E) Pull-down experiment by using GST-GBD and GST-GBD^{Q1022A-T1025A-S1081A} from cells synchronized in cytokinesis. (F) Immunofluorescence staining of GFP-GBD and GFP-GBDQ^{1022A-T1025A-S1081A} (green) with g-tubulin (red) during cytokinesis. Enlarged section shows colocalization with centrosome and midbody. If not previously specified, all results are shown as mean or representative picture of at least three independent experiments ± SEM. ***P < 0.001.