FIG. 3.

Western blot analysis of RsbT variants in B. subtilis. Stationary-phase cultures of B. subtilis (BSA661 T₀ to T₁₈), expressing rsbR-S and either wild-type rsbT (wtT) or one of the rsbT variants (i.e., T₁, T₂,…, etc.) from a pDRNT plasmid integrated at sigB were analyzed by Western blotting as described in Materials and Methods with anti-RsbR and anti-RsbT monoclonal antibodies as probes. The positions of the RsbR (R) and RsbT (T) proteins are indicated. The band that migrated above RsbR is a cross-reactive B. subtilis protein (ca. 40 kDa) present in stationary-phase extracts (15). Lane M contains “broad-range” prestained sodium dodecyl sulfate-polyacrylamide gel electrophoresis molecular mass standards (Bio-Rad, Hercules, Calif.) that were electrophoresed and transferred with the crude extracts.