Localization dynamics of actin-YFP and fluorescent PH domains tracking phosphoinositides. (A–D) RP/RC measured the localization of YFP chimeras to phagosomes, relative to free CFP, averaged over multiple events, in wild-type (black circles), and SHIP1−/− macrophages (white circles). Phagocytosis was completed in 3–5 mins. Error bars represent standard error of the mean (SEM). Horizontal bars indicate Student’s t-test for significant differences. Grey bars indicate p-value > 0.05; black bars indicate data with p-value < 0.05. (A) Actin-YFP on the phagosome during phagocytosis (WT n=17, KO n=24). (B) Plcδ1PH-CIT recruitment (WT n=10, KO n=10). (C) BtkPH-CIT recruitment (WT n=10, KO n=8). (D) Tapp1PH-CIT recruitment (WT n=12, KO n=10). (E) BtkPH-CIT and TappPH-CFP on the phagosome during phagocytosis. (WT n=10, KO n=9). The ratio between BtkPH-CIT and TappPH-CFP was measured and plotted as RP/RC.