Figure 4.
Induction of glucosylation of Rac 1 and activation of caspase 3. (A) Vero cells were exposed to different doses of TcdB or CPD mutants. Cells were harvested 4 h later and western blot was performed using monoclonal antibody (Clone 102) that only binds to non-glucosylated Rac1. β-actin was used as an equal loading control. The right panel shows the relative band density of the non-glucosylated Rac1 to β-actin from three independent experiments; (B) Vero cells were exposed to the indicated doses of TcdB or TcdB-C698S for 24 hr. The activation of caspase 3 was assessed by antibody against cleaved caspase-3 (clone 5A1E). β-actin was used as an equal loading control. The right panel shows the relative band density of the non-glucosylated Rac1 to β-actin from three independent experiments (* indicated that P<0.05 between TcdB and TcdB-C698S).