Figure 6
Protein and mRNA levels of Atf1 transcription factor in cells lacking both flp1 and pcr1 genes upon oxidative stress induction. (a,b) Wild-type and ∆flp1 ∆pcr1mutant cells (2∆) were grown in YES medium to mid-log phase and treated with 1 mM H2O2 for the indicated time points. (a) Total protein extracts were resolved by SDS-PAGE and analyzed by Western blot. Atf1 was detected by incubation with a monoclonal anti-Atf1 antibody. Ponceau staining was used as loading control. Full-length blots are shown in Supplementary Fig. S12. (b) Total RNAs were prepared from untreated (0´) and treated cells at the time points indicated. mRNA levels of atf1+ were measured by qPCR, normalized against ribosomal 18S mRNA and determined with respect to time 0´ values, which were considered as 1. Data represent the average of two biological replicates.
