Fig. 2

Phosphorylation analysis of the TORC1 targets under laboratory conditions. a Immunoblot analysis of Rps6 phosphorylation in the different strains treated with TORC1 inhibitors. The selected laboratory (BY4743, BQS252 and CEN.PK) and industrial strains (C9, EC1118 and M2) were grown in SD medium with the required auxotrophic supplements. Part of the exponentially growing cells was harvested as a control, and the remainder was treated with caffeine (20 mM), MSX (2 mM) and Rapamycin (200 ng/mL) for 30 min. Total lysates were analyzed using the specific anti pS235/S236Rps6 antibody, while total Rps6 was used as the loading control. b Rps6 phosphorylation during the transition from a poor nitrogen source (proline) to a good nitrogen source (ammonium or glutamine). The same conditions as in panel a. Alcohol dehydrogenase (Adh1) was used as an additional loading control. c Par32 phosphorylation during nutrient starvation. Strains carrying the epitope Myc-tagged versions of PAR32 were changed from SD to minimal medium without either glucose or ammonium sulfate for 20 min