Fig. 4

Glab inhibits the activation of the cGAS-STING pathway by disrupting the STING-IRF3 interaction. A Transfection of Flag-tagged plasmids (Flag-MAVS, Flag-STING, Flag-TBK1 and Flag-IRF3) into HEK-293 cells for 12 h, and then treated with vehicle, Glab (20 μM) for 6 h. Whole cell lysates were collected and immunoblotted with the indicated antibody. Samples for qPCR were detected by qPCR assay for the expression of IFN-β mRNA. B BMDMs were first treated with DMSO or Glab (20 μM) for 1 h and then stimulated with cGAMP for 2 h. The expression of STING and the oligomerization of SITNG in the cell lysate were analyzed by Western blot with the indicated antibodies. C and D HEK-293 T cells were transfected with Flag-tagged plasmids (Flag-Vector, Flag-IRF3 and Flag-TBK1) and HA-tagged plasmids (HA-Vector and HA-STING) for 20 h, then treated with Glab (20 μM) for 6 h and immunoprecipitated with Anti-FLAG® M2 Affinity Gel, as shown by Western Blots analysis. Data in A are expressed as mean ± SEM (n = 3/group, from three biological replicates.). one-way ANOVA and Dunnett’s post hoc test were used to assess differences among groups, ^*p < 0.05, ^**p < 0.01 and ^***p < 0.001 vs. the control, NS, not significant