Fig. 4: The acute antiviral response is decreased in AICAR-treated cells with Mff phosphorylation.
From: MAVS is energized by Mff which senses mitochondrial metabolism via AMPK for acute antiviral immunity
a WT MEFs were treated with the AMPK activator AICAR (2 mM) for the indicated times. Whole-cell lysates were analyzed by immunoblotting using antibodies against the indicated proteins. The indicated letters (a, b, c) correspond to the endogenous Mff bands observed in WT MEFs. b, c qRT-PCR analyses of IFN-β (b) and IL-6 (c) mRNA in WT MEFs transfected with poly(I:C)-LMW (1 μg/mL) in the absence (−) or presence (+) of AICAR (2 mM, pretreated for 2 h) for the indicated times. d WT MEFs were stained with MitoTracker Deep Red (mitochondria; red) and then treated with or without AICAR (2 mM) for 8 h. After fixation and permeabilization, cells were immunostained with an anti-MAVS antibody (green). Nuclei were stained with Hoechst 33342 (blue). Scale bars: 10 μm (whole) and 2 μm (inset). e, f qRT-PCR analyses of IFN-β (e) and IL-6 (f) mRNA in Mff KO MEFs transfected with poly(I:C)-LMW (1 μg/mL) in the absence (−) or presence (+) of AICAR (2 mM, pretreated for 2 h) for the indicated times. The blue shaded parts of the graphs (0–8 h) represent the acute phase after poly(I:C), and the red shaded parts (8–24 h) represent the tolerant phase in b, c, e, f. Data represent means ± SEM from two independent experiments (n = 4, P values; two-tailed unpaired t test) in b, c, e, f. Immunoblot analyses and all microscopic analyses were repeated independently more than three times with similar results. Source data are provided as a Source data file.
