Irf3 antibody

Cells were lysed with IP lysis buffer (25 mM Tris, pH7.4; 150 mM NaCl; 1% NP-40, 1 mM EDTA, 5% glycerol; protease and phosphatase inhibitors). A total of 300 μg lysates and 1 μg antibodies were used in each immunoprecipitation. IRF3 was immunoprecipitated with the IRF3 antibody (Cell Signaling, 11904) or control rabbit IgG (Millipore, pp64), and the precipitated immune complex was detected with IRF3, p-IRF3 (S396), TBK1 and p-TBK1 (S172) antibodies.

Cells were washed with PBS buffer and fixed with 4% formalin. Then cells were permeabilized with 0.5% Triton X-100. After cells were washed with PBS, they were blocked and stained with primary antibodies, followed by staining with an Alexa Fluor 488 secondary antibody⁴⁵ . Nuclei were stained with DAPI (Sigma). The antibodies used in this research were: IRF3 antibody from Cell Signaling technology (1:200, Cat# 11904); STAT1 antibody from Cell Signaling technology (1:400, Cat# 14994); P-STAT1 antibody from Cell Signaling technology (1:400, Cat# 9167); Calnexin antibody from Cell Signaling technology (1:50, Cat# 2679); GolgiB1 antibody from Sigma-Aldrich (1:500, Cat# HPA011008). Fluorescence images were obtained and analyzed using a laser scanning confocal microscope (Leica TCS SP5).

Cells were washed with PBS and fixed with 4% paraformaldehyde. Then, the cells were permeabilized with 0.5% Triton X-100. After the cells were washed with PBS, they were blocked and stained with primary antibodies before being stained with Alexa Fluor 488-, 594- and 647-conjugated secondary antibodies.^{41 (link)} Nuclei were stained with DAPI (Sigma). The antibodies used in this research were an IRF3 antibody from Cell Signaling Technology (1:200, Cat# 11904), an HA antibody from Sigma-Aldrich (1:100, Cat# H9658), and a Myc antibody from Sigma-Aldrich (1:200, Cat# M4439). Fluorescence images were obtained and analyzed using a laser scanning confocal microscope (Zeiss LSM 800).