Figure 5.

Role of APPL1 in CXCL12-instigated Akt phosphorylation. A, CXCL12-, insulin-, and EGF-instigated Akt phosphorylation was examined in cells depleted of APPL1. Serum-starved HeLa cells transfected with control siRNA (siLuc) or two distinct siRNAs against APPL1 (siAPPL1-1 and siAPPL1-2) were treated with CXCL12 (10 nm), insulin (50 nm), or EGF (100 ng/ml) for 5 min at 37 °C. Whole-cell lysates were analyzed by immunoblotting for pAkt-Ser⁴⁷³, total Akt-1/2, and APPL1. B, immunoblots were analyzed by densitometry. Bars represent the mean of the relative levels of pAkt-Ser⁴⁷³ to siLuc- and CXCL12-treated cells. Error bars represent the S.D. Data were analyzed by two-way ANOVA followed by Tukey's multiple comparison test. The p values for insulin-treated cells are indicated. CXCL12- or EGF-treated samples were not significant. Veh, vehicle.