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. 2014 May 3;42(11):7084–7095. doi: 10.1093/nar/gku373

Figure 1.

Figure 1.

Analysis of Rad53 checkpoint kinase activation in pkc1 mutant strains. Activation of the checkpoint kinase Rad53 was determined by western analysis as slower migrating bands using an anti-Rad53 antibody. The ponceau staining of the membrane is shown as loading control. (A) Exponentially growing cultures of the wild-type (W303-1a) and pkc1ts (JC6-3a) strains were split and incubated for three hours at 25º or 37º followed by 1 h incubation in the absence or presence of 0.2 M HU, 0.04% MMS or 5 μg/ml phleomycin, or were irradiated with 50 J/m2 UV radiation. (B) Exponentially growing cultures of the pkc1ts (JC6-3a) strain transformed with a centromeric plasmid containing the PKC1 gene or an empty vector were assayed as described in (A). (C) Exponentially growing cultures of the tetO7:PKC1 (JCY1471) strain incubated for 8 h in the absence or presence of 5 μg/ml doxycycline and the wild-type (W303-1a) and pkc1Δ (GPY1115) strains grown in the presence of 1 M sorbitol were incubated for 1 h in the absence or presence of 0.2 M HU or 0.04% MMS as indicated.