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. Author manuscript; available in PMC: 2012 May 1.
Published in final edited form as: Mol Microbiol. 2011 Mar 16;80(3):798–810. doi: 10.1111/j.1365-2958.2011.07609.x

Figure 5. Multiple isoforms are detected in crude cell extracts using anti-RsbRA or anti-RsbRB antibody.

Figure 5

(A & B) For IEF in the ranges indicated on the gel images, 50 μg of protein from wild type (PB2) cells isolated before (control, leftmost lane C) or at various times after exposure to heat (50°C, 54°C), ethanol (4%, 10%) or sodium chloride (4%, 8%) were loaded into each lane. Samples from unstressed rsbRA (AΔ1; PB427), rsbRB (BΔ1; PB545), or rsbT (TΔ1; PB421) null mutants were also analyzed (lane C on the right). Western blots were probed with anti-RsbRA (A) or anti-RsbRB (B); gel images are oriented with alkaline regions uppermost and with isoform positions (0, 1, 2) indicated on the left. (C) Blot using anti-RsbRA antibody to probe extracts of wild type (PB2) and rsbRA T205A mutant (PB505) cells isolated before (control, leftmost lane C) or at various times after exposure to 10% ethanol; isoform positions (0, 1, 2) indicated on the left. (D) Quantification of RsbRA isoform signals from three independent ethanol stress experiments. Isoforms (0-P, 1-P, 2-P) are shown as % of total RsbRA detected.