Journal: Molecules and Cells

Article Title: STING Negatively Regulates Double-Stranded DNA-Activated JAK1-STAT1 Signaling via SHP-1/2 in B Cells

doi: 10.14348/molcells.2015.2359

Figure Lengend Snippet: dsDNA directly triggers the activation of JAK1-STAT1 signaling. (A) BJAB cells were tansfected with poly(dA:dT) (2 μg/ml), poly(dG:dC) (2 μg/ml) or ISD (2 μg/ml) for 6 h and the expression levels of IFIT1, IFI44, MX1 and OAS1 were detected by qPCR. (B) BJAB cells were tansfected with ISRE-luciferase reporter for 36 h and then tansfected with poly(dA:dT), poly(dG:dC) or ISD for 18 h. Luciferase assay analysis of the luciferase reporter gene activity. (C) BJAB cells were tansfected with poly(dA:dT), poly(dG:dC) or ISD for 15 min and 30 min. The phosphorylation of JAK1 and STAT1 were detected by Western blot. (D) BJAB cells were treated with JAK1 inhibitor Baricitinib or STAT1 inhibitor Fludarabine for 1 h prior to tansfection with poly(dA:dT), poly(dG:dC) or ISD for 6 h. The expression levels of IFIT1, IFI44, MX1 and OAS1 were detected by qPCR. The data shown represent the means of three independent experiments, and the error bars represent the s.e.m. * p < 0.05, ** p < 0.01, *** p < 0.001; ns denotes p > 0.05.

Article Snippet: BJAB cells were co-transfected with ISRE luciferase reporter plasimd (2 μg, Promega, USA) and renilla luciferase reporter plasmids (0.2 μg, Promega, USA) for 24 h and then transfected with poly(dA:dT), poly(dG:dC) or ISD.

Techniques: Activation Assay, Expressing, Luciferase, Activity Assay, Phospho-proteomics, Western Blot