Anti phosphorylated irf 7

The culture medium consisted of RPMI 1640 and F12 (Gibco-BRL, Life Technologies Corporation, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS, Hyclone, Thermo Fisher Scientific Inc, Waltham, MA, USA). Recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) and IL-4 were purchased from R&D (Minneapolis, MN, USA). The inhibitors for NF-κB activation, including Bay11-7082 and PDTC, were purchased from Calbiochem (Merck KGaA, Darmstadt, Germany) and Sigma (St. Louis, MO, USA), respectively. The fluorescent-conjugated antibodies (Abs) recognizing CD80, CD86, CD83 and HLA-DR were purchased from BD Pharmingen (BD Biosciences, CA, USA). Anti-CCR7, recombinant IFN-λ1 and chemoattractants, including CCL19 and CCL21, were purchased from R&D Systems. Anti-Flag antibody was purchased from Sigma. Anti-NS1 antibody was purchased from Genetax (GeneTex Inc, Irvine, CA, USA). Anti-phosphorylated IRF-3 and anti-phosphorylated IRF-7 were purchased from Cell Signaling (Beverly, MA, USA). Antibodies recognizing total IRF-1, total IRF-3, total IRF-7, IFN-λR1, COX-2 and upstream transcription factor2 (USF2) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Protein expression was evaluated by Western blot as described before^{37 (link)}. The following antibodies were used for immunoblotting: anti-rabbit and anti-mouse horseradish peroxidase-conjugated secondary antibodies (1:5000; Pierce); anti-human Hsp90 (1:1000; 610418, BD Biosciences), anti-ADAR1 (1:1000; 14175; Cell Signaling); anti-MDA5 (1:500; 5321; Cell Signaling); anti-RIG-I (1:1000; 3743; Cell Signaling); anti-phosphoSTAT1 (1:1000; 9167; Cell Signaling); anti-phosphorylated IRF7 (1:1000; 12390; Cell Signaling); anti-GAPDH (1:1000; ab9485; abcam); and anti-HPV16 E7 (1:1000; ab82601; Abcam).