Immunoblots

Total protein lysates were isolated in lysis buffer (20 mmol/L Tris base, 1% Triton X‐100, 50 mmol/L NaCl, 250 mmol/L sucrose, 50 mmol/L NaF, 5 mmol/L Na₂P₂O₇, plus Halt™ protease and phosphatase inhibitor cocktail [Thermo Scientific]). Equal amounts of protein (100 μg) were loaded and resolved on 10% SDS polyacrylamide gels and transferred to nitrocellulose membranes. Blots were probed with antibodies against ACSL1 (catalog No. 4047), phosphorylated p70 (P‐p70) S6 kinase (S6K) (catalog No. 9234), phosphorylated 4E‐BP1 (catalog No. 2855), and phosphorylated AMP‐activated protein kinase (P‐AMPK) (catalog No. 2531), and were then stripped and reprobed with p70 S6K (catalog No. 9092), 4E‐BP1 (catalog No. 9644), or AMPKα (catalog No. 2532) antibodies, respectively (all antibodies from Cell Signaling). GAPDH (Abcam, catalog No. ab8245) was used as loading control.