RPM protects hepatocytes via the mTORC2-Akt pathway, (a) mTOR inhibitors regulate hepatocyte mTORC2-Akt signaling pathway. Hepa1 cells were incubated with vehicle (Ctl) or RPM, or Torin 1 or Tg, or RPM/Tg, or Torin 1/Tg, as described in the material and methods. Cell lysates were collected at 6h, and analyzed by Western blots to measure levels of phosphorylated S6K, Akt (S473) and LC3B. 2 replicates/expt. group, (b) Akt inhibition abrogated cytoprotective effect of RPM. Hepa1 cells were incubated with vehicle (Ctl) or Tg, or RPM, or RPM/Tg, or TCN or RPM/TCN/Tg, as described in the material and methods. Cell death was measured at 24h by LDH assay. 3 replicates/expt. group.
Representative results of 2 different experiments. *p<0.05, **p<0.01