HOMESEARCHCONTACT US
ResearchPersonnelPublications

Activated Signal Transduction Kinases Frequently Occupy Target Genes

Study Design

Data Insights Supplemental Material
Acknowledgements

Strains

Strains used in this study were originally generated by the J. Weissman lab (Ghaemmaghami et al. 2003; Huh et al. 2003) unless otherwise indicated. The proper tagging of proteins was confirmed by Western blot analysis and by PCR of genomic regions. For gene deletions, the URA3 marker from the plasmid pRS306 was amplified by PCR using primers that are homologous to regions of the gene to be deleted and transformed into yeast. Stable integration of the marker into the gene of interest was selected on URA3- plates and confirmed by genomic PCR analysis.


Growth Conditions

  • As default, yeast cells were grown at 30 °C in rich medium containing 2% glucose (YPD). Biological replicates were grown as independent cultures.
  • Osmotic stress was induced by addition of NaCl (0.4 M final concentration) for 5 minutes.
  • Pheromone induction was performed by addition of alpha pheromone 5ug/ml) for 5 minutes.
  • For growth in a non-fermentable carbon source, complete synthetic medium supplemented with 3% glycerol was used.
  • Oxidative stress was induced by adding hydrogen peroxide (0.4 mM final concentration) and incubation for 20 minutes.
  • Galactose induction was performed by adding galactose (2% final concentration) to a cell culture grown in raffinose-containing medium and incubation for 1 hour.

    •  
    COLLABORATORSINTERNAL SITEQUICK LINKS
       
    YOUNG LAB
    Whitehead Institute
    9 Cambridge Center
    Cambridge, MA 02142
    [T] 617.258.5218
    [F] 617.258.0376
    CONTACT US