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The Prr1 response regulator is not important for Spc1 activation after oxidative stress. Wild-type (KS1376), Δprr1 (CA425), Δmcs4 (CA220), and Δmcs4 Δprr1 (CA423) strains carrying the spc1:HA6H allele were grown to midlog phase at 30°C in YES medium and treated with oxidative stress induced by 0.3 mM H2O2. Aliquots of cells were harvested at the indicated times, and Spc1 was purified by Ni-NTA chromatography, followed by immunoblotting with anti-phospho-p38 and anti-HA antibodies.
