Figure S5.
Effect of NCLX inhibition on CCCP- and oligomycin-activated glycolysis. (A) ECAR (± SD) upon DMSO, 100 µM CK666, 80 µM CGP37157, 1 µM CCCP, 1 µM CCCP + 100 µM CK666 addition, or 1 µM CCCP + 80 µM CGP37157 addition (at 23 min, pink arrow), followed by 50 mM 2-DG (at 89 min, blue arrow) in 2 mM glucose medium without serum. n = 8 individual well measurements per condition. (B) ECAR (± SD) upon DMSO, 100 µM CK666, 80 µM CGP37157, 1.5 µM oligomycin, 1 µM oligomycin + 100 µM CK666 addition or 1 µM oligomycin + 80 µM CGP37157 addition (at 23 min, pink arrow), followed by 50 mM 2-DG (at 89 min, blue arrow) in 2 mM glucose medium without serum. n = 8 individual well measurements per condition. (C) OCR (± SD) in MEFs (in 2 mM glucose without serum) upon 100 µM CK666, 1 µM CCCP or 1 µM CCCP + CK666 addition at 15 min, then 50 mM 2-DG at 59 min. n = 3 individual well measurements for CCCP and CK666; 4 for CCCP + CK666. Pink arrow indicates drug treatment and blue arrow indicates 2-DG treatment. (D) OCR (± SD) in MEFs (in 2 mM glucose without serum) upon DMSO, 100 µM CK666, 2.5 µM antimycin A or 2.5 µM antimycin A + 100 µM CK666 addition at 33 min, then 50 mM 2-DG at 258 min. n = 5 individual well measurements per condition. Pink arrow indicates drug treatment and blue arrow indicates 2-DG treatment. (E) OCR (± SD) in MEFs (in 2 mM glucose without serum) upon DMSO, 100 µM CK666, 2.5 µM rotenone or 2.5 µM rotenone + 100 µM CK666 addition at 33 min, then 50 mM 2-DG at 258 min. n = 5 individual well measurements per condition. Pink arrow indicates drug treatment and blue arrow indicates 2-DG treatment. Number of experiments and sample sizes are provided in Table S1.

Effect of NCLX inhibition on CCCP- and oligomycin-activated glycolysis. (A) ECAR (± SD) upon DMSO, 100 µM CK666, 80 µM CGP37157, 1 µM CCCP, 1 µM CCCP + 100 µM CK666 addition, or 1 µM CCCP + 80 µM CGP37157 addition (at 23 min, pink arrow), followed by 50 mM 2-DG (at 89 min, blue arrow) in 2 mM glucose medium without serum. n = 8 individual well measurements per condition. (B) ECAR (± SD) upon DMSO, 100 µM CK666, 80 µM CGP37157, 1.5 µM oligomycin, 1 µM oligomycin + 100 µM CK666 addition or 1 µM oligomycin + 80 µM CGP37157 addition (at 23 min, pink arrow), followed by 50 mM 2-DG (at 89 min, blue arrow) in 2 mM glucose medium without serum. n = 8 individual well measurements per condition. (C) OCR (± SD) in MEFs (in 2 mM glucose without serum) upon 100 µM CK666, 1 µM CCCP or 1 µM CCCP + CK666 addition at 15 min, then 50 mM 2-DG at 59 min. n = 3 individual well measurements for CCCP and CK666; 4 for CCCP + CK666. Pink arrow indicates drug treatment and blue arrow indicates 2-DG treatment. (D) OCR (± SD) in MEFs (in 2 mM glucose without serum) upon DMSO, 100 µM CK666, 2.5 µM antimycin A or 2.5 µM antimycin A + 100 µM CK666 addition at 33 min, then 50 mM 2-DG at 258 min. n = 5 individual well measurements per condition. Pink arrow indicates drug treatment and blue arrow indicates 2-DG treatment. (E) OCR (± SD) in MEFs (in 2 mM glucose without serum) upon DMSO, 100 µM CK666, 2.5 µM rotenone or 2.5 µM rotenone + 100 µM CK666 addition at 33 min, then 50 mM 2-DG at 258 min. n = 5 individual well measurements per condition. Pink arrow indicates drug treatment and blue arrow indicates 2-DG treatment. Number of experiments and sample sizes are provided in Table S1.

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