Research Paper Volume 12, Issue 2 pp 1114—1127

Figure 4. The overexpression of Romo1 triggered the metabolic reprogramming in macrophages. (A) The levels of IL-10, TGF-β, TNF-α and IL-6 were analyzed by ELISA in control and Romo1-expressed macrophages. (B) The levels of iNOS were analyzed by flow cytometry in control and Romo1-overexpressed macrophages. (C) The extracellular acidification rates (ECAR) of control and Romo1-overexpressed macrophages were measured under basal conditions followed by the sequential addition of 10nM glucose (Glu), 0.5μM oligomycin (Oli) and 100mM 2-deoxyglucose (2-DG). (D) The oxygen consumption rates (OCR) of control and Romo1-overexpressed macrophages were measured under basal conditions followed by the sequential addition of 0.5μM oligomycin (Oli), 1μM carbonyl cyanide p-trifluoromethoxy-phenylhydrazone (FCCP) and 1μM rotenone and antimycin A (R+A). (E) The levels of 2-NBDG were analyzed by flow cytometry in control and Romo1-overexpressed macrophages. The MFIs of 2-NBDG were quantified and statistically analyzed. (F) The mRNA levels of Glut1 and Glut3 were determined by qRT-PCR in control and Romo1-overexpressed macrophages. (G) The relative ATP levels of the control and Romo1-overexpressed macrophages. Data are representative of at least three independent experiments and are presented as mean ± SD. ns, not significant; *, P < 0.05; **, P < 0.01.