Intestinal calcium and bile salts facilitate germination of Clostridium difficile spores
WT and Δ32980 spores were incubated with 0.2% Tc and the indicated concentrations of glycine (a-b) or CaCl2 (c-d) with germination was measured by loss of OD600 at 37°C over the course of an hour. Western blot assessing SleC activation from WT and Δ32980 spores incubated at 37°C for 15 minutes with 1% Tc and the indicated concentrations of glycine (e) or CaCl2 (f). %pro-SleC and % SleC relative densities were calculated for each lane using ImageJ by determining the ratio of each band to the total density for the two bands combined. Blots are representative of 3 experiments. Germination assays were performed in triplicate and are representative of 3 independent spore preps. Error bars are mean plus or minus SD.