Research Paper Volume 12, Issue 2 pp 1114—1127

Romo1 is involved in the immune response of glioblastoma by regulating the function of macrophages

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.