Fig. 3.

CBB-stained polyacrylamide gel (A) and Western blot analyses (B, C) of mutants and their parental strains. (A) Samples (1.2 µg protein) prepared as described under Materials and methods were applied to the 5–20% gradient SDS-polyacrylamide gel. (B) Diluted samples (12 ng protein) were applied to the gel for the following Western blotting analysis. Anti-PAc (SpaP) serum diluted 1:2000 was used as the first antibody. The ECL Plus Western blotting reagents (GE Healthcare, Buckinghamshire, UK) were used for detection. (C) Samples for Z1, E9UA2, and E5UA1 (1.2 µg) as well as for UA159, E4UA1, and E4Z1 (12 ng) were applied to the gel and determined by Western blotting analysis.