Macrophages were lysed with Pierce IP lysis buffer (Thermo Fisher Scientific) with cOmplete Mini Protease Inhibitor and Phosphatase Inhibitor (Roche). After brief sonication on ice, samples were centrifuged at 10,000g at 4°C for 10 minutes and supernatants were saved for Western blotting. Next, 5–30 μg protein was loaded per well onto a NuPAGE 4% to 12% Bis-Tris Gel (Life Technologies). After protein transfer, membranes were incubated with 1:1000 dilution of specific antibodies against phospho-STAT1 (Ser727, 9177), phospho-STAT1 (Tyr701, 9167), STAT1 (14994), and IRF1 (8478) obtained from Cell Signaling Technology. Membranes were incubated with 1:2000 dilution of antibody against β-actin (4970) obtained from Cell Signaling Technology or 1:1000 dilution of antibody against α-tubulin (ab4074) obtained from Abcam as loading control. Antibodies against BACH1 (HRP conjugated, sc-271211) and HO-1 (HRP conjugated, sc-390991) were obtained from Santa Cruz Biotechnology and utilized at 1:1000 dilutions.

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