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. Author manuscript; available in PMC: 2013 Nov 1.
Published in final edited form as: Mol Microbiol. 2012 Sep 27;86(3):645–660. doi: 10.1111/mmi.12006

Fig. 5.

Fig. 5

Representative Western blots of Phos-tag gels containing extracts of walK and pnpRS mutant strains grown exponentially in BHI broth. Growths, rapid preparation of protein extracts, 75 μM Phos-tag SDS-PAGE, and quantitative Western blotting with anti-WalRSpn antibody are described in Experimental procedures and Supplemental Information. The positions of unphosphorylated WalR, WalR~P, and a very faint contaminant band are indicated. (A) walK+ parent strain (IU1781); ΔwalK (IU1896); walKΔPAS (IU2306); walKH218A (IU3102); walKT222A (IU5401); and control heated sample of walKT222A (IU5401) showing heat lability of WalR~P. (B) walK+ ΔpnpR (IU3483); walK+ ΔpnpRS (IU4086); ΔwalK ΔpnpR (IU5720); ΔwalK ΔpnpRS (IU5728); and control heated sample of ΔwalK ΔpnpR (IU5720). Additional heated controls for extracts of other strains are shown in Fig. S12. The faint contaminant band was at the limit of detection and still present in extracts of a ΔwalR Pc-pcsB+ strain (EL1472), which lacks WalR and WalR~P (see Fig S11).