Extended Data Fig. 1: Activation of Fgr is necessary for M1-like macrophage polarization.
From: Fgr kinase is required for proinflammatory macrophage activation during diet-induced obesity
a, BMDM from WT and FgrKO mice untreated or treated with LPS + IFNγ were analyzed by flow cytometry for NO2, IL12p40, TNFα, CD86 and CD40 (n = 6). b-c, Measure of H2O2 (b) and superoxide (c) in WT BMDM treated under the conditions indicated (n = 11). d, Pyruvate-malate and (e) succinate driven respiration in isolated liver mitochondria in the indicated treatments (n = 6). f, Maximal respiratory capacity of mitochondrial complex I, II and IV in isolated liver mitochondria in the indicated treatments (n = 6). g, BMDM from WT and FgrKO mice untreated or treated with xanthine/xanthine oxidase (X/XO), LPS, the combination of both or adipose conditioned media (AT) were analyzed by flow cytometry for NOS2, MHC-II, NOS2 and CD86. WT, n = 6 for all conditions but 33% AT where n = 3, FgrKO, n = 3. Representative unnormalized seahorse profile of oxygen consumption of BMDM from WT and FgrKO mice treated with LPS+ IFNγ for 16 h (overnight, o/n, h) or 4 h (i) (n = 7 for untreated or n = 4 for treated). Representative unnormalized oxygen consumption analysis of IFNγ primed (pIFNγ) BMDM from WT (left panels) and FgrKO (right panels) treated with (j) LPS, (k) LPS plus NPA and (l) LPS plus NAC at the start of the SeaHorse assay (n = 3). m, Complex II immunocapture and phosphorylation analysis by western blot of SDHA in WT BMDM untreated or treated with LPS+ IFNγ for 4hrs. (a,g) *, P <0.05; **, P <0.01; ***, P <0.001; ****, P <0.0001. 2-way ANOVA and Tukey post-hoc test was used. Each point represents a biological replicate. Data are the mean ± SEM.
