Fig. 2. Protein levels during a pheromone time course. (A) Western blot of total cell extracts from a pheromone time course probed with anti-Tea1, anti-GFP (for Tea2GFP) and anti-Tip1 antibodies. New forms of protein appearing after pheromone addition are marked by arrows. (B) Cells were shifted to 36°C in the presence or in the absence of pheromone for 4 hours. Cells at 36°C in the presence of pheromone arrest in G1 but do not activate the shmooing growth mode (Fig. 7B,C). Cells were then released at 25°C to allow synchronous progression into shmooing growth. Samples were taken at time 0, after 4 hours at 36°C and after 1 hour release and western blots were carried out on total cell extracts for Tea1p. (C) Native cell extracts were made from vegetatively growing cells and treated with -phosphatase in the absence () or in the presence (+inh) of phosphatase inhibitors. An untreated sample is run as a control (C). Western blots were probed for Tea1p. The lower dephosphorylated form, which increases after phosphatase treatment, is marked by an arrow.