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. 2017 Feb 10;8:14391. doi: 10.1038/ncomms14391

Figure 7. IFI16 regulates cGAMP-mediated STING activation.

Figure 7

(a) Control, IFI16, cGAS, STING KO THP-1 cells or (b) MDMs with IFI16 siRNA knockdown, were infused with cGAMP (50 nM) at indicated time-points and subsequently evaluated for type I interferon secretion. (c) STING dimerization analysis by semi-native western blotting. Upper lane represents an overexposure of the dimer STING band. Total STING was run on a separate SDS–Page gel. (d) Control and IFI16 KO cells were infused with cGAMP (50 nM) for 2 h, fixed and stained for DAPI (blue), IFI16 (green) and IRF3 (red). (e) IRF3 translocation from cytoplasm to nuclear saturation were quantified by counting >50 separate images of control or IFI16 KO cells 2 h post cGAMP infusion. (f) Subcellular fractions of control and IFI16 KO cells stimulated with 50 nM cGAMP for 1 h were immunoblotted for phosphorylated IRF3 and total IRF3 in cytosolic (cyto) and nuclear (nucl) fractions. Data in (a,b) represent mean±s.d. of biological triplicates from (a) three independent experimental setups or (b) one donor; (c–f) data is representative of one of three independent experiments.