Figure 2.

Metabolic restriction with MYX alters ROS and ATP production in human monocytes in the absence of glucose. (A–C) Oxygen consumption rate (OCR) and inhibitor sensitive oxygen consumption rate (ΔOCR) of quiescent and stimulated (100 ng/mL LPS) human monocytes after 2 h (A1-C1) 6 h (A2-C2) of incubation in the presence or absence of Myxothiazol (MYX) (pmol/10⁶ cells) or 1% (v/v) DMSO (vehicle control) under glucose free conditions. After preincubation, (A) basal OCR, (B) VAS2870 sensitive OCR (ΔOCRV) and (C) oligomycin sensitive OCR (ΔOCRO) were assessed (mean ± SEM; n = 9 – 10). (D) Intracellular ATP content of quiescent and stimulated (100 ng/mL LPS) human monocytes after 3 h of incubation in the presence or absence of MYX (pmol/10⁶ cells) or 1% (v/v) DMSO (vehicle control) under glucose free conditions was measured by bioluminescence (mean ± SEM; n = 6). (E) Median fluorescence intensity of quiescent and stimulated (100 ng/mL LPS) human monocytes after 2 h of incubation in the presence or absence of MYX (pmol/10⁶ cells) or 1% (v/v) DMSO (vehicle control) and/or 50 µM VAS2870, or 100ng/ml PMA or H₂O₂ under glucose free conditions, stained with CM-H₂DCFDA (mean ± SEM; n = 3 – 5); *p < 0.05, **p < 0.01, ***p < 0.001. Wilcoxon signed rank test. ^#p < 0.05; Mann-Whitney U Test.