Fig 7. Tor1 N-terminal HEAT repeats were required for adequate cell wall- and heat stress responses.
A, E. Dilutions of cells of indicated genotypes were spotted on YPD medium without or with 300 ng/ml doxycycline (300 Doxy), cell wall stress was induced with 10 ng/ml micafungin (Mica) or 15 μg/ml Congo red (CR) (A), and heat stress was induced at 43o (E); strains TOR1/TOR1, JKC1361 for YPD and micafungin and JKC1713 for Congo red and heat stress; tor1/TOR1, JKC1345, JKC1346, JKC1347; tetO-TOR1-Del381, JKC1442, JKC1445, JKC1441; tetO-TOR1, JKC1543, JKC1546, JKC1549). B. Cells of indicated genotypes were grown in YPD medium containing Vehicle (H2O) or 15 ng/ml micafungin (Mica), without or with 300 ng/ml doxycycline (left panel); or in synthetic complete medium (SC) containing Vehicle (H2O) or 4 μM nikkomycin (Nikko), without or with 300 ng/ml doxycycline (right panel). Strains TOR1/TOR1, JKC1713; tor1/TOR1, JKC1347; tetO-TOR1-Del381, JKC1441; tetO-TOR1, JKC1549. Error bars show SD of 3 technical replicates. C, D. Cells of indicated genotypes were pre-grown in YPD medium with 5 ng/ml doxycycline for 3.5 h (Time 0) and then diluted into fresh YPD medium with 5 ng/ml doxycycline with either 10 ng/ml micafungin (Mica) or H2O as vehicle (Veh, D lower panel is the same P-S6 blot as Fig 1B). Total protein extract was probed with antibody to phosphorylated Mkc1 (P-Mkc1) and the PSTAIRE antigen of Cdc28 as loading control (C), or with antibody to phosphorylated Rps6 (P-S6) and tubulin (Tub) as loading control (D). Dens: signal intensity ratio of P-Mkc1 to Cdc28 (C) or P-S6 to Tub (D); strains TOR1/TOR1, JKC1713; tetO-TOR1-Del381, JKC1441; tetO-TOR, JKC1549. Representative of 2 biological replicates.