Inverse correlation between TORC1 and PKC1activities. A) The PKC1 pathway was activated in wt cells upon inhibition of TORC1 with rapamycin. This pathway is constitutively activated in myo1Δ cells. A–B) All histograms show the ratio of the intensities of each P-Slt2p band relative to the intensity of its Pgk1p loading control, averaged from duplicate experiments. Error bars represent STDError mean. B) Steady state levels of hyper phosphorylated Slt2p (P-Slt2p, 55 kDa) were assayed by Western blot using equal amounts of protein extract (50 μg) from myo1Δ, tor1Δ, and myo1Δtor1Δ strains treated with rapamycin (+) or with DMSO alone (-). Pgk1p was used as a loading control. C) Limiting dilution growth assay on agar medium measuring relative viability of wt, myo1Δ, tor1Δ, and myo1Δtor1Δ strains. 10-fold dilutions are indicated at the top of the image (see Methods for details).
Pagán-Mercado et al. BMC Cell Biology 2012 13:13 doi:10.1186/1471-2121-13-13