Supplementary Figure 6: FAMIN-deficient M2 macrophages exhibit impaired mitochondrial ROS- and FAO-dependent production of extracellular ROS.
From: C13orf31 (FAMIN) is a central regulator of immunometabolic function
(a,b) Intracellular ROS measurement in unstimulated M1 and M2 mFamin–/– and mFamin+/+ macrophages (MΦ) stained with the cytosolic ROS indicator, 5-(and-6)-chloromethyl-2-7-dichlorodihydrofluorescein diacetate, acetyl ester (CM-H2DCFDA) and measured in relative fluorescence units, RFU. (c–i) Zymosan induced eROS production in M1 and M2 macrophages treated as indicated for 16 h with 20 μM C75; or 1 h 40 μM etomoxir (ETO) or 500 μM mitoTEMPO prior to stimulation; or silenced for Fasn (Fasn siRNA), Cpt1a (Cpt siRNA) or Cybb (Cybb siRNA) or scrambled siRNA (Ctrl siRNA); Left, eROS kinetic plots measured in relative light units, RLU and right, area under curve, AUC. (j) NADPH quantification in M1 and M2 macrophages cell lysates. (k) PMA-induced eROS production in mFamin–/– and mFamin+/+ neutrophils. *P < 0.05, **P < 0.01 (Unpaired, two-tailed Student’s t-test). Data are from one experiment with three mice representative of two independent experiments (a–k; mean ± S.E.M.).
