Galactose allows for selective interference with aerobic glycolysis and inhibits key aspects of activated M1 macrophages, which show metabolic plasticity.
(a) Real-time measurement of ECAR of BMDMs, unstimulated (control) or stimulated with IFN-γ after incubation in medium containing glucose, galactose or glucose +10 mM 2-DG for 1 h, vertical line indicates time of injection of IFN-γ. Data are representative of three independent experiments (n = 4–5, mean ± sem).
(b) Real-time measurement of OCR of BMDMs, unstimulated (control) or stimulated with IFN-γ after incubation in medium containing glucose, galactose or glucose +10 mM 2-DG for 1 h, vertical line indicates time of injection of IFN-γ. Data are representative of three independent experiments (n = 4–5, mean ± sem).
(c) ECAR of BMDMs, unstimulated (control) or stimulated with IFN-γ after incubation in medium containing glucose or galactose for 1 h. *p < 0.05, ***p < 0.001, NS: no significant difference. Data are representative of two independent experiments (n = 3–4, mean ± sem).
(d) OCR of BMDMs, unstimulated (control) or stimulated with IFN-γ after incubation in medium containing glucose or galactose for 1 h. *p < 0.05, ***p < 0.001, NS: no significant difference. Data are representative of two independent experiments (n = 3–4, mean ± sem).
(e) OCR/ECAR ratio of BMDMs, unstimulated (control) or stimulated with IFN-γ after incubation in medium containing glucose or galactose for 1 h. *p < 0.05, ***p < 0.001, NS: no significant difference. Data are representative of two independent experiments (n = 3–4, mean ± sem).
(f) Cell viability of BMDMs after stimulation with IFN-γ in medium containing glucose or galactose ± oligomycin (1 μM) at the indicated time intervals. Data are representative of two independent experiments (n = 6, mean ± sem).
(g) BMDMs were stimulated with IFN-γ for 24 h in medium containing glucose or galactose. ATP/ADP ratio was tested after treatment with DMSO or oligomycin for 30 min. ***p < 0.001, NS: no significant difference. Data are representative of two independent experiments (n = 4, mean ± sem).