Extended data Figure 2. Measuring mitochondrial membrane potential in vitro in A549 and L3161C cells.

a, Gating strategy used for quantification of TMRE signal. R2 – region representing single cells was used for quantification of the TMRE signal. b, Overlay histogram showing shifts in TMRE staining in L3161C cells strained with Vehicle, 8 μM Oligomycin, and 8 μM Oligomycin + 4 μM FCCP. c, TMRE measurements in A549 cells treated with indicated concentrations of Phenformin or FCCP for 3 hr (n = 3 biological replicates). d, TMRE measurements in mouse cell line L3161C treated with indicated concentrations of Phenformin or FCCP for 3 hr (n = 3 biological replicates). e, Viability of A549 cells treated with indicated concentrations of Phenformin for 3 hr (n = 3 biological replicates). f, Uptake of ¹⁸FBnTP probe measured by gamma counter in A549 cells treated with 1 mM Phenformin for 3 hr (n = 5 biological replicates). g, Oxygen consumption rate (OCR) per cell measured in A549 cells treated acutely with 1 mM Phenformin (n = 25 technical replicates). h, OCR per cell measured in mouse cell line L3161C treated acutely with 1 mM Phenformin (n = 25 technical replicates). i, TMRE measurements in mouse cell line L3161C treated with Vehicle, 8 μM Oligomycin, or 8 μM Oligomycin with 4 μM FCCP for 3 hr (n = 3 biological replicates). j, Uptake of ¹⁸FBnTP probe measured by gamma counter in mouse cell line L3161C treated with Vehicle, 8 μM Oligomycin, or 8 μM Oligomycin with 4 μM FCCP for 3 hr (n = 6 biological replicates). k, Viability of L3161C cells treated as in j (n = 6 biological replicates). Data are shown as mean +/− SD. Experiments in c-i, were repeated twice with similar results. Experiments in j and k were done once.