FIG 2.

Generation of specific gene knockout cell lines by CRISPR/Cas9-mediated genome editing. Knockout gene variants in HFFs for cGAS (cGAS KO), STING (STING KO), and IFI16 (IFI16 KO), were generated using CRISPR-Cas9 technology. The efficiency of IFI16, cGAS, and STING protein depletion was assayed by RT-qPCR for cGAS, STING, IFI16, and the GADPH housekeeping gene (the data are shown as mean fold changes plus SD; ***, P < 0.001 by two-way ANOVA followed by Bonferroni's posttest for comparison of KO versus WT cells) (A); by Western blot analysis for cGAS, STING, IFI16, and vinculin as a loading control (B); and by TIDE analysis to quantify indel frequencies and composition using PCR amplicons spanning the sgRNA target sites for Sanger sequencing and subsequent analysis with TIDE software (http://tide.nki.nl) (C and D).