Fig. 2

Nutrient effects on kinase activities and developmental induction. a Cell growth rates in full growth media without (Med) or with 500 nM rapamycin (Med+Rap). Doubling times represent mean ± standard error from triplicates in an independent experiment (N = 3). b p4EBP1 and pAMPKα of adhered cells in full growth media in the presence and absence of 500 nM rapamycin. Log-phase cells were adhered to plate wells in fresh full growth media. After 2 h, fresh media were replenished with or without 500 nM rapamycin. Samples were taken at various times and analyzed by immunoblotting. Nutrient withdrawal (DB), but not rapamycin, induced expression of developmental genes and multi-cell formation. Developmental protein expression (c) and aggregation (d) were examined in adhered cells upon nutrient removal (DB) or in full growth media containing rapamycin. c Immunoblots represent three independent experiments. e Log-phase cells in full growth media were transferred to indicated strengths of diluted full growth media and growth rates measured. After 24 h, immunoblotting was performed and represent three independent experiments. f Log-phase cells in full growth media were transferred to indicated strengths of glucose-free (Glc^[-]) media, supplemented with varying concentrations of glucose. Growth rates were then measured. After 24 h, samples were analyzed by immunoblotting. Immunoblots are representative of three independent experiments. g Adhered log-phase cells in DB or Glc^[-] media containing indicated concentrations of glucose. After 5 h, samples analyzed by immunoblotting. Immunoblots represent three independent experiments. h Adhered log-phase cells in DB with or without 80 mM glucose and/or rapamycin as indicated. At various times, samples were analyzed by immunoblotting. Immunoblots represent three independent experiments. i Adhered log-phase cells in DB with or without mM glucose and/or rapamycin as indicated. After 5 h, cells were visualized for developmental aggregation