Vinculin knock-down in 293 cells does not affect the uptake of S. aureus. (A) 293 cells were stably transduced with pLKO.1 or pLKO.1 encoding shRNA against human vinculin (pLKO.1 sh-hvinc). Whole cell lysates were prepared and WCLs were analyzed by Western blotting for vinculin, FAK, paxillin, Src, and β-tubulin expression. Representative blots are shown from 2 independent experiments. (B and C) Cells from (A) were seeded on gelatine coated 24 well plates and infected with S. aureus at MOI 20 for 2 h. Total cell associated or recovered viable intracellular bacteria were determined by bacterial cell adhesion (B) or gentamicin protection assays (C), respectively. Values are means ± SEM from 4 independent experiments done in triplicate, n=12. Data were analyzed for statistical significance using Mann-Whitney test.