Fig 2: Reprogramming macrophage metabolism into that of M1 macrophages. Mouse bone marrow-derived macrophages (BMDMs) were harvested and treated with IL-4 (10 ng mL−1) for 12 h to induce M2 macrophage switching. A The levels of intracellular glycogen in M2 macrophages treated with CHIR99021:CH (3 μM), SB415286:SB (3 μM) or NaW (2 mM) were measured by colorimetric assay. B The protein levels of phosphorylated GSK3β-9Ser, GSK3β, p-Gys1 and Gys1 were measured by Western blotting. C, D The expression of Ugp2 and Pygl in M2 macrophages with or without NaW treatment was analyzed by real-time PCR (C) and (D) western blotting. E, F Liquid chromatography–tandem mass spectroscopy (LC‒MS/MS) was performed for mearing UDPG (E) and G6P/G1P (F) levels in M2 macrophages treated with or without NaW. G–J The expression of Arg1, Tgfβ1, Il10, Tnf, Il6 and Nos2 in M2 macrophages with or without NaW treatment was analyzed by real-time PCR (G). iNOS, Arg-1, TGFβ, IL-10, TNF and IL-6 levels were measured by western blotting (H) and ELISAs (I, J). K‒M M2 macrophages were pretreated with a glycogen phosphorylase inhibitor (GPI) for 6 h. The expression levels of Arg1, Tgfβ1, Il10, Tnf, Il6 and Nos2 in M2 macrophages treated with or without NaW were measured by real-time PCR (K), IL-10, TGFβ, IL-6 and TNF expression was measured by ELISAs (L), and iNOS protein expression was measured by western blotting (M). N The expression of Arg1, Tgfβ1, Il10, Tnf, Nos2 and Il6 in M2 BMDMs pretransfected with siRNA (Pygl) and treated with or without NaW before IL-4 stimulation as measured by real-time PCR. O, P The expression of P2Y14 in M2 macrophages treated with or without NaW was measured by real-time PCR (O), and the expression of STAT1 and p-STAT1 was measured by western blotting (P). Unless otherwise specified, n = 3 biologically independent experiments. The data are presented as the mean ± SEM. P values were calculated by one-way ANOVA. *P < 0.05; **P < 0.01; ***P < 0.001