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. 2018 Apr 20;9(30):21166–21181. doi: 10.18632/oncotarget.24989

Figure 3. Metformin reduces oxygen consumption rate of lymphoma cells.

Figure 3

Daudi cells (A, B) and SUDHL-4 (C, D) were pretreated with 0 μM (Black), 100 μM (white), 1 mM (grey) metformin for 7 days and their metabolic state was determined using the Seahorse XF Cell Mito Stress test (Agilent). In short, after basal measurements, cells were stressed with 1 μM oligomycin (inhibits ATP synthase) first and OCR measurements were taken and then 0.5 μM FCCP for Daudi and 1 μM FCCP for SUDHL-4 cells (depolarizes mitochondrial inner membrane) was added and OCR measurements were taken and finally Antimycin A and rotenone were added together to inhibit mitochondrial respiration and OCR was measured again. No metformin was present during the assay (2 hours). Arrows in (A) and (C) represent addition of each stress compound. Basal respiration = (Last OCR measurement before oligomycin injection) – (nonmitochondrial respiration rate after antimycin A and rotenone injection). Spare respiratory capacity = (Maximal respiration after FCCP addition)-(Basal respiration). p values for basal OCR and spare respiratory capacity at 0 μM vs 1 mM metformin is shown. *****p value < 0.0001; **p value < 0.005; ns = not significant.