Fig 5.
Truncation of RsiV results in constitutive degradation in a RasP-dependent manner. Immunoblot comparing full-length RsiV from B. subtilis Phs-gfp-rsiV (JLH453) and Phs-gfp-rsiV ΔrasP::cat (JLH466) strains and RsiV without the extracellular domain from Phs-gfp-rsiV1-60 (JLH516) and Phs-gfp-rsiV1-60 ΔrasP::cat (JLH536) strains. WT strains (JLH453 and JLH516) were subcultured 1:100 into 5 ml LB supplemented with IPTG (0.5 mM). Strains containing rasP mutations (JLH466 and JLH536) were subcultured 1:50 into 5 ml LB supplemented with IPTG (0.5 mM). Cultures were grown to mid-log phase and then left untreated (−) or treated (+) with lysozyme (2 μg/ml) for 10 min at 37°C. The immunoblot was probed with anti-GFP antibodies or anti-σA antibody, which was used as a loading control. Cytoplasmic GFP was used as a size control (TPM1263).