HSV-induced Akt phosphorylation dependent on αvβ3-integrin, its C-tail, and TLR2.

<p>(A, B, D–F) Akt phosphorylation requires β3-integrin and TLR2. wt or sh-β3 cells 293T (A, B), HaCaT (D), HeLa (E), SK-N-SH (F) cells were transfected or not with plasmid encoding TLR2-Flag, silenced for β3-integrin as indicated (sh-β3), exposed to R7910 (60 PFU/cell) for 10, 20 and 30 min. Cells were lysed and total proteins were subjected to SDS-PAGE. Phosphorylated and total Akt were detected with PAb to phospho-Akt (Ser473) (P-Akt) or PAb to total Akt (T-Akt). (C) β3-integrin silenced (sh-β3) 293T were transfected with wt-β3-integrin or β3_Y747-759F plus αv-integrin, and TLR2. Cells were cultured in pre-exhausted medium <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004477#ppat.1004477-Gianni2" target="_blank">[8]</a> for 48–72 h from 12 after transfection, and then exposed to R7910 (60 PFU/cell) for 10, 20, 30 min, or uninfected (0′). Total cell lysates were subjected to SDS-PAGE and WB for P-Akt or T-Akt.