Anti phospho stat2

Dual luciferase assay, confocal immunofluorescence microscopy, immunoprecipitation and Western blotting were performed as previously described [50 (link)–52 (link)]. Relative luciferase activity in arbitrary units was calculated by normalizing firefly luciferase activity with Renilla luciferase activity.
Mouse anti-c-myc (clone 9E10) was purchased from MilliporeSigma. Mouse anti-V5 and anti-GST were from Thermo Fisher Scientific. Rabbit anti-STAT1, rabbit anti-STAT-2, mouse anti-JAK1 (A-9), and mouse anti-GAPDH were purchased from Santa Cruz Biotechnology (TX, USA). Rabbit polyclonal antibodies against phospho-STAT1, phospho-JAK1 were bought from Cell Signaling Technology (MA, USA). Anti-phospho-STAT2 was purchased from R&D Systems (MN, USA).

Monoclonal antibodies (Abs), specific for cluster of differentiation (CD)1a, CD14, CD38, CD86, CD83, HLA-DR, CD40, IgG1, and IgG2a (BD Bioscience, San Diego, CA, USA), were directly conjugated to fluorescein isothiocyanate (FITC) or phycoerythrin (PE). To exclude dead cells from the analysis, Fixable Viability Dye eFluor®780 (FvDye) (eBioscience, San Diego, CA, USA) was used. For immunoblotting analysis, rabbit anti-STING (Cell Signaling, Danvers, MA, USA # 2775), anti-IRF3 (Santa Cruz, Santa Cruz, TX, USA # sc-9082), anti-IRF7 (Santa Cruz, # sc-9083), anti-STAT1 (BD Bioscience, # 610186), anti-phospho STAT1 (Cell Signaling Technology, Leiden, The Netherlands, # 7649), anti-STAT2 (BD Transduction Laboratories, # 610188), anti-phospho STAT2 (R&D Systems, Minneapolis, MN, USA, MAB2890), mouse anti-actin (Sigma-Aldrich, St. Louis, MO, USA #A0483), and horseradish peroxidase-conjugated secondary antibody anti mouse (Santa Cruz, # sc-2005) and anti rabbit (Santa Cruz, # sc-2004) were used. For phagocytosis and phagosomal acidification experiments, cytochalasin D 5 μM (Sigma-Aldrich, # C8723) and chloroquine 2 μM (Sigma-Aldrich, # C6628) were used.