Western Blot Analysis of Immune Signaling

Cells and decidual tissues were lysed in RIPA buffer (Thermo Scientific, Massachusetts, USA) with protease and phosphatase inhibitor (Thermo Scientific, Massachusetts, USA) and analyzed by western blot as described previously 31 (link). Protein concentration was determined by BCA protein assays kit (Thermo Scientific, Massachusetts, USA) according to the manufacturer's instruction. Antibodies against STAT1 (14995, Cell Signaling Technology, Danvers, MA, USA), phosphorylated STAT1 (p-STAT1, 9167, Cell Signaling Technology, Danvers, MA, USA), inducible nitric oxide synthase (iNOS, 2982, Cell Signaling Technology, Danvers, MA, USA), IRF1 (8478, Cell Signaling Technology, Danvers, MA, USA), interferon regulatory factor-5 (IRF5, 20261, Cell Signaling Technology, Danvers, MA, USA), interferon regulatory factor-8 (IRF8, 5628, Cell Signaling Technology, Danvers, MA, USA) and GAPDH (ab181603, Abcam, Cambridge, MA, USA) were used according to the manufacturer's instruction.