Figure 3.

Cytotoxicity of TcdBs in HeLa cells. (A) HeLa cells were treated with 100 pM of TcdB_VPI (VPI), TcdB_NAP1 (NAP1), and TcdB_NAP1v (NAP1v) for 24 h. Cytotoxicity was analyzed by flow cytometry using propidium iodide (PI)/anexin V double staining. Control cells were left untreated (mock). Error bars represent means ± SD of three independent experiments; (B) Following the addition of 100 pM of TcdB, the cells were lysed at the indicated time points, and the dynamics of Rac1 glucosylation was monitored by immunoblot with a specific anti-Rac1 antibody that only recognizes the unmodified form of this protein (ungluc.Rac1). Untreated cells (-) were included as a positive control for unglucosylated Rac1, and immunodetection of beta-actin served as a loading control (β-actin). Shown are representative western blot images from three independent experiments.