Fig 1: ASK1 phosphorylates IRF3.a, b Representative western blot and quantification of pIRF3 protein levels in isolated adipocytes of HFD-fed ASK1F/F and ASK1Δadipo mice. n = 6 mice per group. *p = 0.048. c, d Representative western blot and quantification of pIRF3 upon incubation of recombinant IRF3 with or without active ASK1. n = 6 biological replicates. ***p = 0.0006. e, f Western blot and quantification of pIRF3 protein levels in subcutaneous adipocytes transfected with lentivirus expressing wild-type (pLV-CMV-ASK1; ASK1 wt) or kinase-negative (pLV-CMV-ASK1-K716R; ASK1 mut) ASK1. n = 3 biological replicates. #p = 0.070. g, h Representative western blot and quantification of pIRF3 protein levels in subcutaneous adipocytes transfected with control siRNA (Co siRNA) or siRNA targeting ASK1 (ASK1 siRNA) treated with or without 100 ng/ml LPS for 6 h. n = 4 biological replicates. ****p < 0.0001. i Ucp1 mRNA expression in subcutaneous adipocytes transfected with control shRNA lentivirus (shLuc) or shRNA lentivirus targeting IRF3 (shIRF3) pre-treated with 100 ng/ml LPS for 24 h followed by stimulation with 0.1 µM isoproterenol for 6 h. n = 6 biological replicates. *p = 0.019. Values are expressed as mean ± SEM. Statistical tests used: two-sided t-tests for (b, d, f, i), two-sided one sample t-test for (h). Source data are provided as a Source Data file.