Fig. 1. GS enhanced mitochondrial respiration capacity and ATP production in cardiomyocytes and neurons. (A-B) Basal oxygen consumption rate (OCR) in H9c2, NGF-induced PC12, neuron, HUVECs and BMSCs treated with different concentrations of total ginsenosides (GS) for 48 h was measured using a MitoXpress® Xtra probe normalized to cell number. (C)OCR was analyzed at basal condition and sequential injections of oligomycin (1 µM), FCCP (5 μM), rotenone (Rot, 1 μM) and antimycin A (AA, 1 μM) in control and GS-pretreated H9c2 cells by the MitoStress Test Kit. After the normalization to cell number, basal OCR, ATP production, maximal respiratory capacity (MRC), and spare respiratory capacity (SRC) were calculated and shown. (D) Similar to the panel C, basal OCR, ATP production, MRC, and SRC were analyzed in NGF-induced PC12 cells, after GS pretreatment for 48 h. (E) After pretreatment with GS at 2.5, 5, or 10 μg/mL for 48 h, ATP levels in H9c2, PC12, and neurons were measured by the Bioluminescence Detection Kit. (F)The intensities of ATP and ADP in control and GS-treated H9c2 cells were analyzed by the LC-MS/MS method based on MRM detection. The ratio of ATP and ADP was calculated and shown. (G-H) Basal OCR and ATP content in H9c2 cells treated with different concentrations (2.5, 5, or 10 μg/mL) of GS and incubated for 3, 6, 12, 24, or 48 h were measured as described as the methods above. Ctrl: control group. *P < 0.05, **P< 0.01, and ***P < 0.001 versus Ctrl group.