Figure 3.

Autophagy inhibition reduces glycolysis and induces OXPHOS. ECAR (A) and OCR (B) were measured in shATG7-expressing cells using the Seahorse Extracellular Flux Analyzer. Glycolysis (relative to shCtrl-expressing cells) was calculated as “average ECAR following glucose addition minus average ECAR following inhibition of glycolysis using 2-DG.” Relative mitochondrial respiration was calculated as “basal OCR minus OCR following antimycin and rotenone (ETC inhibitors) treatment (legends for Fig. S3 provide more details). Three independent experiments were performed in quintuplicate. ECAR (C) and OCR (D) were measured in K562 cells following 24 h 10 µM HCQ treatments. Two independent experiments were performed in quintuplicate. (E and F) ShATG7-expressing K562 cells were cultured in the absence (E) or presence (F) of U-¹³C₆ for 24 h. Following cell lysis, intercellular levels and incorporation of labeled carbons in ATP was measured by LC-MS. Two independent experiments were performed in triplicate. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001. +2, etc. corresponds to ATP containing 2X, etc. ¹³C.