Protein Extraction and Immunodetection

Protein extraction, quantification, and immunodetection were performed as in [16 , 21 (link)]. Membranes were incubated O/N with the corresponding primary antibody dilutions: mouse anti-β-ACTIN (1:5000, A5441, Sigma-Aldrich, San Luis, Missouri, USA), goat horseradish peroxidase (HRP)-conjugated anti-GAPDH (1:3500, sc-365,062, Santa Cruz, Dallas, Texas), rabbit anti-ATG4A (1:1000, #7613, Cell Signaling, Danvers, Massachusetts, USA), rabbit anti-LC3B (1:3000, 51,520, Abcam, Cambridge, UK), rabbit anti-MSI2 antibody (1:1000, EP1305Y, Abcam, Cambridge, UK), mouse anti-P62 (1:1000, 65,416, Abcam, Cambridge, UK), mouse anti-P21 (1:2000, #2946, Cell Signaling, Danvers, Massachusetts, USA), mouse anti-SCD1 (1:1000, 19,862, Abcam, Cambridge, UK). After three washes with 1x PBS-T, membranes were incubated for 1 h at RT with the corresponding secondary antibody dilutions: goat HRP-conjugated anti-rabbit-IgG (1:3500, A0545, Sigma-Aldrich, San Luis, Missouri, USA) or goat HRP-conjugated anti-mouse-IgG (1:5000, B7264, Sigma-Aldrich, San Luis, Missouri, USA). Images were acquired with an ImageQuant LAS 4000 or AMERSHAM ImageQuant 800 (GE Healthcare) and were quantified using ImageJ software (NIH).