Fig 1: G10 activity is STING variant dependent.G10 activates the luciferase coupled ISRE.ISG54 reporter system in R232, H232, HAQ variants of human STING to varying degrees in stably transfected HEK293T cells. Cells were incubated with G10 for 18 hours and luciferase activity was estimated.
Fig 2: G10 does not promote phosphorylation of HAQ STING or IRF3 in THP-1 cells.Cells were incubated with G10 for 2 hours and then proteins harvested and analyzed by immunoblotting using human pSTING, STING, pIRF3 and actin antibodies. ADU-S100 was used as a positive control.
Fig 3: Murine STING pathway proteins are not phosphorylated by G10 treatment.Murine STING and IRF3 in CT26 cells that stably overexpress mSTING are not phosphorylated after G10 treatment. Cells were incubated with G10 for 2 hours and then proteins harvested and analyzed by immunoblotting using murine pSTING and STING and pIRF3 antibodies, with DMXAA (mouse specific) and ADU-S100 (pan STING) as positive controls.
Fig 4: Compound 11 is a potent STING agonist in the cell free pSTING assay.Compound 11 is a direct STING agonist. Immunoblots showing Compound 11 dependent phosphorylation of recombinant CTT domain of STING by TBK1 (Lane 1: No agonist, Lanes 2, 3: Compound 11 at 10 and 5μM respectively).
Fig 5: G10 promotes phosphorylation of STING pathway proteins in THP1-R232 cells.G10 treatment leads to phosphorylation of the R232 variant of human STING in THP1 knock in cells. Cells were incubated with G10 for 2 hours and then proteins harvested and analyzed by immunoblotting using human pSTING, STING, pIRF3 and actin antibodies. ADU-S100 was used as a positive control.
Supplier Page from Cayman Chemical for STING R232 variant (human, recombinant)