Figure 1

rSTING and hSTING respond in a more similar way to anticancer/antiviral agents DMXAA or CMA than the way of mSTING.

(A) HEK293T cells were transiently transfected with indicated species of STING plasmids, along with IFN β-Luc reporter plasmids. After 12 hours, cells were PFO-permeabilized to deliver cGAMP linkage isomers (5 μM) or stimulated with DMXAA (266 μM) or CMA (500 μg/mL). Luciferase reporter assay was performed after incubation for an additional 12 hours. Values represent the mean average of triplicate experiments. Error bars indicate SEM. (B) Similar to (A), 293T cells were transfected with indicated species or mutated STING plasmids and stimulated with 2′3′-cGAMP, DMXAA or CMA. Immunoblot analysis (with anti-p-IRF3, anti-STING and anti-GAPDH) was performed after 12 hours. GAPDH as the loading control. The assays were run under the same experimental conditions and the blots were cropped from original full-sized images (attached as supplemental Fig. S8). (C) RT-PCR analysis of indicated STING-pathway related gene expression levels after indicated stimulation in THP-1 (human monocytic cells) or peritoneal macrophages from rat or mouse. HPRT as the loading control. Full RT-PCR are shown in Supplemental Fig. S8. (D) ITC binding curves for titration of DMXAA or CMA into indicated species of STING-CTD.