FIG. 3.

RV39 increases phosphorylation of Ser⁴⁷³Akt in a Src- and PI 3-kinase-dependent manner. (A) 16HBE14o− cells without or with inhibitor (1 h at 37°C) were treated with sham or RV39 infection (MOI of 1, 10 min at 33°C). Detergent-soluble proteins (500 μg) were immunoprecipitated with mouse monoclonal 4G10 anti-phosphotyrosine (2 μg). Immunoprecipitates were immunoblotted for phospho-Tyr⁴¹⁶ (p-Tyr⁴¹⁶) Src (top panel) or total Src. (B) Group mean data for three experiments (means ± standard errors of the means; asterisk indicates a value different from that for RV alone; P < 0.05 by ANOVA). (C) Whole-cell lysates were immunoblotted for phospho-Ser⁴⁷³ Akt (20 μg protein/lane). The PP2 Src family kinase inhibitor blocked phosphorylation of both Src and Akt more effectively than PP3 did. These results are representative of three experiments.