Figure 3. Concentration-dependent stimulation of EGFP-LC3 processing and inhibition of mTORC1 signaling by perhexiline, niclosamide, amiodarone and rottlerin.

MCF-7 cells stably expressing EGFP-LC3 were treated for 4 h with chemicals at the indicated concentrations or with 20 nM rapamycin. EGFP-LC3 processing was monitored by probing lysates with anti-GFP antibody, mTORC1 activity by probing lysates with antisera against phosphorylated (Thr³⁸⁹) S6K and total S6K, and mTORC2 activity with antisera against phosphorylated (Ser⁴⁷³) PKB/Akt and total PKB/Akt. β-tubulin immunostaining was used as a protein loading control.