Fig. 2. Tor2 promotes cell growth and is part of the TORC1 complex. (A) Wild-type (tor2+) and tor2-51 cells were grown in EMM to mid-log phase at 25°C, shifted to the restrictive temperature of 32°C, and samples were taken at the indicated times. RNA was extracted and northern blot performed and hybridised using probes against the ribosomal protein genes rps1102 and rpl1701. The rRNA in each sample, stained with Methylene Blue, is shown to verify equal loading of RNA in each lane. (B,C) Crude extracts were prepared from cells exponentially growing in rich medium. The cells expressed tagged proteins, Mip1-myc, Ste20-myc or Pop3-myc, and HA-Tor1 or HA-Tor2, as indicated. The extracts were immunoprecipitated (IP) with the indicated antibodies (, anti-) and the precipitates were subjected to SDS-PAGE and examined by western blot (WB). Immunoblots of the crude extracts are also shown.