Extended Data Fig. 2: Measuring mitochondrial membrane potential in vitro in A549 and L3161C cells.

a, Gating strategy used for the quantification of TMRE signal. The R2 region representing single cells was used for quantification of the TMRE signal. b, Overlay histogram showing shifts in TMRE staining in L3161C cells treated with vehicle, 8 μM oligomycin or 8 μM oligomycin plus 4 μM FCCP. c, TMRE measurements in A549 cells treated with the indicated concentrations of phenformin or FCCP for 3 h (n = 3 biological replicates). d, TMRE measurements in mouse cell line L3161C treated with the indicated concentrations of phenformin or FCCP for 3 h (n = 3 biological replicates). e, Viability of A549 cells treated with the indicated concentrations of phenformin for 3 h (n = 3 biological replicates). f, Uptake of ¹⁸F-BnTP probe measured by gamma counter in A549 cells treated with 1 mM phenformin for 3 h (n = 5 biological replicates). g, 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 h (n = 3 biological replicates). j, Uptake of ¹⁸F-BnTP probe measured by gamma counter in mouse L3161C cells treated with vehicle, 8 μM oligomycin, or 8 μM oligomycin with 4 μM FCCP for 3 h (n = 6 biological replicates). k, Viability of L3161C cells treated as in j (n = 6 biological replicates). Data are mean ± s.d. Experiments in c–i, were repeated twice with similar results. Experiments in j and k were done once.