Supplementary Figure 4: cGAS associates with TTLL4 and TTLL6.

(a, b) WT BMDMs co-transfected with CCP5 (a) or CCP6 (b) and cGAS-wt or cGAS-mut for 24 h were immunoprecipitated with antibody against Flag, followed by immunoblotting with the indicated antibodies. (c, d) Ccp5^−/− (c) and Ccp6^−/− (d) BMDMs transfected with cGAS-wt or cGAS-mut for 24 h were immunoprecipitated with antibody against Flag, followed by immunoblotting with the indicated antibodies. (e, f) GST-tagged CCP5-mut (e) or CCP6-mut (f) was incubated with lysates from cells overexpressed with WT or mutant cGAS for 24 h, followed by a GST pulldown assay. (g) BMDMs, DCs and lung fibroblasts from WT mice were subjected to RNA extraction and RT-PCR analysis of the indicated genes. (h) GST-tagged WT (FL) or truncated cGAS was incubated with BMDM lysates for 4 h, followed by a GST pulldown assay. (i, j) GST-tagged wt or mutant cGAS was incubated with recombinant TTLL4 (i) or TTLL6 (j) for 4 h, followed by a GST pulldown assay. (k) Knockout schemes for Ttll4 and Ttll6 knockout mouse generation using CRISPR/Cas9 technology. The targeting exon was shown (upper panel) and the knockout allele was sequenced (lower panel). Data are shown as means±SD. Data are representative of at least three independent experiments.