HeLa cells (1 × 104) were seeded in 96-well plates [33 (link)]. The culture medium was removed the next day, and 100 µL of C-CM, N-CM, or H-CM were applied to the cells. After 12, 24, or 48 h of incubation, the cells were stained using an Orange Mitochondrial Membrane Potential Assay Kit (Abcam, Cambridge, UK). The fluorescence ratio (Ex/Em = 540/590 nm) indicating MMP was measured using a Mithras2 LB 943 Multimode Reader (Berthold Biotechnologies, Bad Wildbad, Germany) [33 (link)].
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