Supplementary Figure 5: CCP5 or TTLL4 does not affect the DNA-binding ability of cGAS.

(a) Confocal microscopy of Ccp5^+/+ and Ccp5^−/− BMDMs transfected with FITC-conjugated 50-mer dsDNA (left panel). Pearson’s Correlation Coefficient between cGAS and FITC-DNA was calculated (right panel). Colocalized dots were annotated by white arrow heads. Scale bar, 10 μm. (b) ChIP assay of 200-mer dsDNA transfected CCP5^+/+ and CCP5^−/− BMDMs expressing WT or mutant cGAS. (c) ChIP assay of 200-mer dsDNA transfected Ttll4^+/+ and Ttll4^−/− BMDMs expressing WT or mutant cGAS. (d) In vitro DNA pulldown of WT or mutant cGAS glutamylated by TTLL4. (e) Ttll6^+/+ and Ttll6^−/− BMDMs were incubated with HSV-GFP (MOI=1) for 24 h, followed by microscopy examination. Cells were counterstained with DAPI for nucleus (left panel). GFP positive cells were calculated (right panel). Scale bar, 400 μm. (f) Cgas^−/− BMDMs rescued with cGAS-wt or cGAS-E272A were incubated with HSV (MOI=1) for 8 h, followed by detection of Ifnb. (g) Cgas^−/− BMDMs rescued with cGAS-wt or cGAS-E272A were incubated with HSV-GFP (MOI=1) for 24 h, followed by microscopy examination. Cells were counterstained with DAPI for nucleus (left panel). GFP positive cells were calculated (right panel). Scale bar, 400 μm. Data are shown as means±SD. ∗, P<0.01; **, P<0.001. Data are representative of at least four independent experiments.