Figure 2 | Scientific Reports

Figure 2

From: A new negative feedback mechanism for MAPK pathway inactivation through Srk1 MAPKAP kinase

Figure 2

MAP phosphatase levels and activity are not dependent on Srk1. (a) and (b), S. pombe wild-type, srk1∆, and wis1∆ cells expressing genomic pyp2:HA6his (a) and pyp1:HA6his (b) were grown in YES medium to mid-log phase and remained untreated (0 min) or treated with 1 M KCl at the indicated times, up to 60 min. Activated (Sty1-P) and total Sty1 were detected with anti-phospho p38 and anti-HOG1 antibodies, respectively. Total Pyp2 (a) and Pyp1 (b) levels were detected with anti-HA antibody. *longer exposure of Pyp2 and P-Sty1 levels are shown in Δwis1. (c) Wild-type, pyp1∆ and pyp2∆ cells with ( +) or without (-) Srk1 overexpression (OP Srk1: through the inducible plasmid pREP1-srk1:HA6his) during 20 h, were untreated (0 min) or treated with 1 M KCl for 15 min. Activated (Sty1-P) and total Sty1 were detected with anti-phospho p38 and anti-HOG1 antibodies, respectively. The level of overexpressed Srk1 was detected with anti-HA antibody. All blot membranes were cut prior antibodies incubation, (d) Decimal dilutions of strains of the indicated phenotypes were spotted on YES solid plates with or without 1 M KCl that had been incubated at 30 °C for 3 days and photographed.

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