Fig. 2. Inhibition of mGPDH by iGP-1 increases OXPHOS and glycolysis in HDMB03 cells (Group 3 MB).

HDMB03 (A) and D-458 (C) cells were seeded at the same density and incubated in 21%, 6%, and 1% O₂ for 24 h and 72 h in the absence (Ctl) or presence of iGP-1 (1 or 100 µM). Cell proliferation was measured using an ADAM cell counter. HDMB03 (B) and D-458 (D) cells were seeded at the same density and incubated in 21%, 6%, and 1% O₂ for 24 h and 72 h in the absence (Ctl) or presence of iGP-1 (1 or 100 µM). Cell viability was measured using an ADAM cell counter. A–D The 2-way ANOVA is representative of at least three independent experiments. Not significant (ns), *p < 0.05 and **p < 0.005. Respiratory control of HDMB03 cells. OCR was measured in real-time with the XF96 analyzer. Cells were cultured for 24 h in Nx (21% O₂ - E) and Hx (1% O₂ - F) in the absence (Ctl) or presence of iGP-1 (1 or 100 µM). Cells were deprived of glucose for 1 h, then glucose (G), oligomycin (O), DNP, and Rotenone + Antimycin A (R/A) were injected at the indicated times. The graphs are representative of at least three independent experiments carried out in octuplicate. Respiratory control of D-458 cells. OCR was measured in real time with the XF96 analyzer. Cells were cultured for 24 h in Nx (21% O₂ - G) and Hx (1% O₂ - H) in the absence (Ctl) or presence of iGP-1 (1 or 100 µM). Cells were deprived of glucose for 1 h, then glucose (G), oligomycin (O), DNP, and Rotenone + Antimycin A (R/A) were injected at the indicated times. The graphs are representative of at least three independent experiments carried out in octuplicate. ECAR of HDMB03 cells in Nx (21% O₂ - I) and Hx (1% O₂ - J) in the absence (Ctl) or presence of iGP-1 (1 or 100 µM) for 24 h was evaluated with the XF96 analyzer. Cells were deprived of glucose for 1 h, then glucose (G) and oligomycin (O) were injected at the indicated times. The graphs are representative of at least three independent experiments carried out in octuplicate. ECAR of D-458 cells in Nx (21% O₂ - K) and Hx (1% O₂ - L) in the absence (Ctl) or presence of iGP-1 (1 or 100 µM) for 24 h was evaluated with the XF96 analyzer. Cells were deprived of glucose for 1 h, then glucose (G) and oligomycin (O) were injected at the indicated times. The graphs are representative of at least three independent experiments carried out in octuplicate. E–L Black star (*) represents the statistical differences between iGP1 (1 and 10 µM) and control, gray star (*) between iGP1 (1 µM) and control and orange (*) star between iGP1 (10 µM) and control. The 2-way ANOVA is representative of at least three independent experiments. *p < 0.05, **p < 0.005, ***p < 0.001 and ****p < 0.0001.