Western Blot Analysis of Cellular Signaling

Western blot analyses were performed as described previously.^{10 (link)} Antibodies used in this study were as follows: phospho-AKT (Ser473; #9271), AKT (#9272), phospho-GSK3β (S9; #9323), LC3B (#3868), phospho-mTOR (S2448; #2971), mTOR (#4517), p-S6 (S235/S236; #4856), S6 (#2217) and β-TrCP (#4394) from Cell Signaling (Danvers, MA, USA); β-TrCP (sc-390629), HSP90 (sc-7947), c-Myc (sc-764) from Santa Cruz (Santa Cruz, CA, USA); cyclin E (51-1459GR) from BD Biosciences (San Jose, CA, USA) and β-actin and horseradish peroxidase-conjugated secondary antibodies from Sigma. Densitometric analysis was performed by ImageJ (NIH, Bethesda, MD, USA).^{12 (link)}