Illumina HiSeq 2500 paired end sequencing; Redox control of the cAMP/PKA pathway during protein misfolding stress
Redox control of the cAMP/PKA pathway during protein misfolding stress
Untreated and treated samples were harvested by centrifugation. Saccharomyces cerevisiae wild-type W303 (MATa ura3-52 leu2-3 leu2-112 trp1-1 ade2-1 his3-11 can1-100) and tsa1D (tsa1::LEU2) were grown in triplicates at 30oC shaking till mid exponential phase in minimal SD medium (0.17% w/v yeast nitrogen base without amino acids with 0.5% w/v ammonium sulfate, 2% w/v glucose supplemented with appropriate amino acids and bases). To induce protein misfolding azetidine-2-carboxylic acid (AZC) (5 mM) was added for two hours.  Untreated and treated samples were harvested and total RNA was isolated using Trizol (Life Technologies, Carlsbad, CA) and quantified using a Nanodrop 8000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA). rRNA was depleted using the RibominusTM Eukaryote Kit for RNA-Seq (Life Technologies, Carlsbad, CA). Truseq stranded mRNA Assay.