Fig. 5: ExcB inhibited STING palmitoylation and signaling.
From: Marine diterpenoid targets STING palmitoylation in mammalian cells
a HEK293T cells expressing wild type (wt) mSTING or indicated cysteine mutants with or without HA tags were labeled with 50 μM alk-16 for 2 h. b HEK293T cells overexpressing mSTING were pretreated with 10 μM of the indicated inhibitors for 1.5 h prior to metabolic labeling with 50 μM alk-16 for 3.5 h. After reaction with azide-biotin, samples before and after NeutrAvidin enrichment were immunoblotted for STING. UT, untransfected. c Levels of total and phosphorylated TBK1 and IRF3 in NIH3T3 cells at indicated times after dsDNA-induced STING signaling. Cells were pretreated with DMSO or 10 μM excB for 1 h prior to dsDNA introduction. d Levels of total and phosphorylated TBK1 and IRF3 in NIH3T3 cells pretreated for 1 h with indicated concentrations of excB followed by dsDNA induction of STING signaling for 2 h. Equal volume of DMSO was used for 0 μM. e THP-1 monocytes containing the IRF-luciferase reporter were pretreated for 2.5 h with indicated concentrations of excB prior to cGAMP addition. IRF reporter activity was normalized to DMSO-treated controls (100%). n = 2.
