FIG 11.

Coimmunoprecipitation (IP) of phosphorylated PI3K and ERK signaling molecules with the V-ATPase pump. (A to D) Serum-starved LLC-PK cells were inoculated with or without the PSaV Cowden strain (MOI of 1 FFU/cell) for the indicated times. Subsequently, the cell lysates were immunoprecipitated using antibodies specific for V₁ subunit E of V-ATPase (A), pPI3K (B), pAkt (C), and pERK (D). The coimmunoprecipitated products were analyzed by Western blotting to detect pPI3K p85 (Tyr458)/p55 (Tyr199), pAkt (Ser473), pERK (Thr202/Tyr204), and V₁ subunit E by use of the relevant antibodies. GAPDH was used as a loading control.