Expression of ROCK-I and ROCK-II was down-regulated in L2 cells by RNAi using shRNA constructs targeting GFP, ROCK-I, ROCK-II or both ROCK-I and ROCK-II. Seventy-two hours post-transfection, L2 cells were infected with wild-type L. monocytogenes (10403S) for 1 h and cell-associated (total) and intracellular bacteria were quantified by gentamicin protection assay. The upper graph indicates the means ±s.d. CFU per well for one of three representative experiments performed in triplicate with similar results. ** p < 0.01 and *** p < 0.001 compared to GFP control. # p < 0.05 compared to ROCK-I and ROCK-II samples. Bottom panels: shRNA-transfected L2 cell lysates were analyzed by Western blot using antibodies specific for ROCK-I, ROCK-II, or tubulin.