Figure 2.

Nur77 Is Required for Normal Mitochondrial Activity in Naive Macrophages

(A and B) Oxygen consumption rate (OCR) after sequential injection of glucose (Glc), oligomycin (OM), carbonyl cyanide 4-trifluoromethoxyphenylhydrazone (FCCP), and antimycin plus rotenone (AA+Rot) in RAW264.7 upon knock down of Nur77 (shNur77) versus control (shCtrl) (A), or in WT and Nur77-KO BMDMs left untreated or stimulated with LPS for 24 hr (B). Bar graphs show OXPHOS parameters derived from OCR values as illustrated in Figure S2C.

(C) Extracellular acidification rate (ECAR) in WT and Nur77-KO BMDMs left untreated or stimulated with LPS for 24 hr. Bar graphs show parameters of glycolysis from ECAR values, as illustrated in Figure S2E.

(D) Relative intracellular ATP in WT and Nur77-KO BMDMs left untreated or stimulated with LPS for 24 hr, followed by vehicle (−) or 5 μM OM (+) for 2 min.

(E–G) Mitochondrial abundance in untreated or 24 hr LPS-stimulated WT and Nur77-KO BMDMs determined by ratio of mitochondrial (mt)DNA to nuclear (n)DNA (E), flow cytometry (fluorescence-activated cell sorting [FACS]) for MitoTracker Green FM staining (F), or live-cell imaging of mitochondria visualized with MitoTracker Green FM (G). MFI, median fluorescence intensity. Micrographs on the left show representative MitoTracker stained cells. Scale bar indicates 20 μm.

Data are shown as means ± SEMs (n = 6 for [A]–[C]; n = 3 for [D], [F], and [G]; n = 106–136 cells for [E]). p values were calculated using two-tailed Student’s t test ([A]–[C], [E]–[G]) or two-way ANOVA with Tukey’s post hoc test (D). n.s., non-significant. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001.

See also Figure S2.