Skip to main content
. Author manuscript; available in PMC: 2013 Oct 23.
Published in final edited form as: Sci Signal. 2013 Apr 23;6(272):ra26. doi: 10.1126/scisignal.2003312

Figure 3. Mating pheromone activates HOG.

Figure 3

A. HOG transcriptional output. Wild-type (LD3342), Δsho1 (RB3704), Δssk1 (RB3382a) and Δssk2 (RB3642) cells adapted to sorbitol were stimulated with α factor and collected into cycloheximide at the indicated time points for imaging. Data corresponds to the mean +/− SEM of the total YFP fluorescence intensity per cell normalized to time zero of one experiment (N = 3 experiments). (n ∼ 700 cells for each data point). YFP fluorescence significantly increased 50 min after pheromone treatment for WT (p < e-4) and Δsho1 (p < e-4), but not for Δssk1 (p = 0.31) or Δssk2 (p = 0.07). B. HOG phosphorylation. Representative WB with protein extracts from LD3342 cells cultured as in A using antibodies against the indicated proteins (left). Quantification of Hog1 phosphorylation in the indicated conditions (right). Data corresponds to the mean +/− SEM of the indicated ratios, normalized to values at time zero (N = 3). Hog1-pp to total Hog1 increases after pheromone treatment (p < e-4). C. HOG localization. Wild-type yeast(ySP69) expressing Hog1-Venus and Hta2-CFP were cultured as in A and imaged in a confocal microscope. Montage showing cells with in-focus nucleus (left). Quantitification of nuclear Hog1-Venus fluorescence localization at the indicated osmolarity (right). Data corresponds to the mean +/− SEM ratio of the ratio of nuclear YFP to CFP signal (N = 3 experiments). Nuclear YFP to CFP ratio increases 50 min after pheromone treatment (p < e-4). D. Model of pheromone-dependent activation of HOG through stimulation of the Sln1 branch. See also Fig. S4 to S7.