Anti mavs

For immunoblotting analysis, lung samples and cell lysates lysed by RIPA buffer (Beyotime, China) were resolved by SDS–PAGE and transferred to the polyvinylidene fluoride membrane (Millipore, United States). Immunoblots were visualized by the ECL system (Fdbio Science, China). The following antibodies were used in immunoblotting analysis: antibodies for anti-IFITM3 (1:2,000), anti-MFN2 (1:2,000), anti-IFN-β (1:500) antibody and anti-IL-1β (1:1,000) were from Abcam. β-actin antibody (1:2,000) was purchased from Fude Biotechnology. Anti-phospho-IRF3 (1:1,000), and anti-TNF-α (1:2,000) antibodies were purchased from Cell Signaling Technology. Anti-MAVS (1:500) antibodies were obtained from Proteintech Group. β-actin was used as an internal control and the relative densities of the measured protein were quantified by image J software.

Primary antibodies to FLAG, GAPDH, CDK1, p-CDK1 (Thr-15), and p-CREB (Ser-133) were purchased from Abclonal Technology. Other antibodies used include anti-MAVS (Proteintech Technology), anti-CREB (Abcam), anti-c-Rel and anti-p-IκBα (S32/35) (Cell Signaling Technology), and anti-mouse/rabbit secondary antibodies labeled with horseradish peroxidase (Boster). The monoclonal antibodies against JEV NS5, NS1 (bind to both NS1 and NS1′), and NS1′ (specifically bind to NS1′) were generated in our laboratory (26 (link)). Poly(I·C) was obtained from Sigma. CDK1 inhibitor RO3306 was purchased from Selleck.