Fig 8.

IEF analysis of phosphorylation of B. subtilis RsbV after exposure to salt stress. B. subtilis PB2 was grown in LB at 37°C with shaking (225 rpm) to the mid-exponential phase (OD₆₀₀, 0.3) and was treated with 64 μM FPSS or an equivalent volume of DMSO. Cultures were then treated with 0.3 M (final concentration) NaCl and were incubated at 37°C with shaking. Samples were removed before (0 min) and after (3 and 8 min) salt treatment. Crude extracts (40 μg protein) were separated using vertical IEF, transferred to nitrocellulose membranes, and probed using a monoclonal anti-RsbV antibody.