Fig. 2.

a Adherence of pilin mutant strains of C. perfringens to C2C12 myoblasts. pilA1 mutant, strain AH7; pilA2 mutant, strain AH8; pilA3 mutant, strain AH9; pilA4 mutant, strain AH10. pAH10 carries a copy of the pilA2 gene under control of a lactose-inducible promoter (5 mM lactose was used to induce pilA2 expression). The mean and SEM of at least five replicates each from at least three independent samples are shown; the P values shown were calculated using the two-tailed students t-test. b Time course showing changes in adherence of strain HN13 to C2C12 myoblasts after transfer from BHI plates to BHI liquid. The mean and SEM of at least five replicates each from two independent samples are shown; the P value shown was calculated using the two-tailed students t-test. c Representative anti-PilA2 Western blots. Lane 1, molecular weight markers with sizes (kDa) indicated on the left; lane 2, strain HN13 grown on BHI plates; lane 3, strain HN13 grown in BHI liquid; lane 4, strain AH8 (ΔpilA2) with pAH10 grown on BHI plates; lane 5, strain AH8 (ΔpilA2) grown on BHI plates with 5 mM lactose. Asterisks denote bands used for densitometry measurements shown in panel D. d Densitometry on four bands identified as specific to PilA2 in Western blots. Images from five individual Western blots were analyzed, and the mean and SEM are shown. “All bands” denotes the total mean pixel intensity for the four bands for each sample that can be most clearly seen in lane 4. The asterisks denote the total mean pixel intensity for the two lower bands visible in panel C that were marked with asterisks