Fig. 3

ROCK mediated LPS-induced ezrin phosphorylation and translocation. A549 and HPAEpiC cells were challenged with LPS (1 µg/mL) in the presence or absence of ROCK siRNA or Y-27632. Total ezrin and p-ezrin protein levels were detected by western blotting using β-actin as an internal reference (a, b). Data are expressed as means ± SD of triplicate samples. *p < 0.05 versus control, #p < 0.05 versus LPS. The intracellular localization of p-ezrin in resting and LPS-activated cells was investigated by immunofluorescence. P-ezrin was stained with Alexa Fluor 488-conjugated IgG (green), F-actin was stained with Rhodamine-phalloidin (yellow), and nuclei were stained with DAPI (blue) (c, d). Qualitative analysis was performed by confocal microscopy. Scale bar, 10 μm. All experiments were performed in three independent experiments