Fig. 4

UBXN3B regulates STING dimerization, phosphorylation, and degradation. a Immunoblots showing Sting dimerization in untreated (Ubxn3b^+/+) and 4-hydroxyl tamoxifen-induced Cre^+/− Ubxn3b^flox/flox (Ubxn3b^−/−) primary MEFs. The cells were infected without (mock) or with HSV-1 (MOI = 0.5) for 8 h. b Immunoblotting analysis of the whole-cell lysates of bone marrow-derived cDCs infected with HSV-1 (MOI = 5). Mono monomer, 2-ME β-mercaptoethanol, a chemical compound that reduces disulfide bonds. c qPCR quantification of Ifnb1 and Tnfa mRNA induction in cDCs by HSV-1 (MOI = 5). Bars: mean ± s.e.m. Two biological replicates were pooled for qPCR (N = 2 per genotype per time point). *P < 0.05 (unpaired Student’s t test). d–f Immunoblotting analysis of the whole-cell lysates of d MEFs infected with HSV-1 (MOI = 0.5), e MEFs transfected with ISD (8 µg/ml) in the absence or presence of 40 µM of chloroquine (+CQ) and f trophoblasts transfected with cGAMP (8 µg/ml). In e, f, the arrows indicate phosphorylated STING with long and short exposure. Actin, Tubulin (Tub), and GAPDH are housekeeping protein controls. g qPCR quantification of IFNB1 and TNFA mRNA induction in trophoblasts transfected with cGAMP (8 µg/ml). In c, g, the bars are: mean ± s.e.m. Two biological replicates were pooled for qPCR (N = 2 per genotype per time point). *P < 0.05 (unpaired Student’s t test). The results are representative of two to three independent experiments