Single-Cell Analysis Reveals That Insulation Maintains Signaling Specificity Between Two Yeast MAPK Pathways with Common Components

Jesse C. Patterson, Evguenia S. Klimenko, and Jeremy Thorner*

*To whom correspondence should be addressed. E-mail: jthorner{at}berkeley.edu

This PDF file includes:

• Materials and Methods
• Results
• Fig. S1. Co-stimulation for 1 hour results in coactivation of the HOG and mating pathway transcriptional reporters in single cells.
• Fig. S2. Co-stimulation for 2 hours results in further coactivation of the HOG and mating pathway transcriptional reporters in single cells.
• Fig. S3. Flow cytometry confirms that dual stimulation leads to dual pathway activation in individual cells.
• Fig. S4. Restricting MAPK activation to the Sln1 branch of the HOG pathway does not alter HOG and mating pathway reporter coexpression.
• Fig. S5. Restricting MAPK activation to the Sho1 branch of the HOG pathway does not alter HOG and mating pathway reporter coexpression.
• Fig. S6. Sonication followed by concurrent stimulation with high concentrations of sorbitol and α-factor causes cell death.
• Fig. S7. Sequential stimulation of cells does not result in mutually exclusive, or any other history-dependent change in, HOG and mating pathway reporter expression.
• Fig. S8. Restricting MAPK activation by the mating pathway to Fus3 does not alter HOG and mating pathway reporter coexpression.
• Fig. S9. Restricting MAPK activation by the mating pathway to Kss1 does not alter HOG and mating pathway reporter coexpression.
• Fig. S10. Both Kss1- and Ste5-dependent Fus3 activation contribute to cross talk in Hog1-deficient cells.
• Fig. S11. Sustained Hog1 catalytic activity is required to prevent cross talk to the mating pathway.
• Fig. S12. Validation of analog-sensitive KSS1 alleles.
• Fig. S13. Lack of Hog1 phosphorylation of Ste50 or Pbs2 does not permit cross talk.
• References

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