Fig. 3

MEKi does not modulate TBK1 activation but upregulates activity of AKT. a Immunoblot analysis (with anti-pTBK1, anti-TBK1, and anti-α-tubulin) of AECs pretreated with DMSO, MEKi, or anti-IFN-β antibody for 1h followed by infection with or without RV2 or RSVA2 or UV inactivated viruses for 24h (MOI 0.1). For the poly (I:C) model (right panel), immunoblot analysis of AECs pretreated with DMSO or MEKi for 1h and subsequently stimulated with or without poly(I:C) for 1-24h. b Immunoblot analysis (with anti-pAKT, anti-AKT, and anti-α-tubulin) of AECs pretreated with DMSO, MEKi, or anti-IFN-β antibody for 1h followed by infection with or without RV2 or RSVA2 or UV inactivated viruses for 24h (MOI 0.1). For the poly(I:C) model (right panel), simultaneous quantification of pAKT / total AKT was determined in AECs pretreated with DMSO or MEKi for 1h and subsequently stimulated with or without poly(I:C) for 1-24h. c Simultaneous quantification of pAKT / total AKT in AECs pretreated with MEKi at the indicated concentration and EC50 determined accordingly. d Simultaneous quantification of pAKT / total AKT in AECs pretreated with PI3Ki at the indicated concentration and IC50 determined accordingly. AlphaLISA analysis of IFN-β in the supernatants of AECs pretreated PI3Ki for 1h followed by stimulation with poly (I:C) for 4h. † relative to MEKi vs untreated and * relative to MEKi/poly(I:C) vs untreated. a and b are representative immunoblots (n = 6). Data are presented as median ± interquartile range in b (poly (I:C) model), c and d. (n = 6). Statistical analysis was performed with the Wilcoxon signed-rank test. * or † p < 0.05 indicates statistical significance