Fig. 5

eCIRP downregulates βPIX and inhibits Rac1 activation. RAW 264.7 cells were incubated with 0.1, 0.2, and 1 µg/ml rmCIRP for 24 h (A) or with 1 µg/ml rmCIRP for 1, 5 and 24 h (B). βPIX levels were assessed by Western blotting. Representative blots are shown. The experiment was performed twice. Data are expressed as the mean ± SD (n = 3–6/group). The PBS control was set to 1 for normalization. The groups were compared by one-way ANOVA and the SNK method. *p < 0.05 vs. PBS. C, D RAW 264.7 cells were incubated with rmCIRP (1 µg/ml) or PBS for 24 h, followed by incubation with or without E. coli for 30 min. Then, GTP-Rac1 levels were evaluated by a GTP-Rac1 pull-down assay. The experiment was performed twice. Data are expressed as the mean ± SD (n = 3/group). The PBS control was set to 1 for normalization. The groups were compared by two-way ANOVA and the SNK method. *p < 0.05 vs. PBS without E. coli; ^#p < 0.05 vs. the PBS-treated group with E. coli. Pos positive control, Neg negative control. E RAW 264.7 cells were incubated with rmCIRP (1 µg/ml) or PBS for 24 h, followed by incubation with or without E. coli for 30 min. Then, F-actin was stained with phalloidin (green), and the nucleus was stained with DAPI (blue). Images were acquired with a confocal microscope, and representative images are presented. Scale bar = 50 µm