Figure 1.

The cGAS-STING pathway is required for increased ISG expression in ASCC3-deficient cells

(A) Schematic of relevant innate immunity signaling pathways.

(B) Western blot analysis of IRF3 and TBK1, and their phosphorylated forms (IRF3ser396 or TBK1ser172), in cells depleted of ASCC3.

(C) qRT-PCR analysis of relative ISG expression in MRC5VA cells treated with siRNAs. Error bars represent standard deviation (SD) of three technical replicates and are representative of three biological replicates.

(D) Western blot analysis of IRF3p (ser396), IRF3, TBK1p (ser172), TBK1, RSAD2, IFIT2, STAT1p (tyr701), STAT1, ASCC3, STING, and MAVS in the same cells as in (C).

(E) Western blot analysis of RSAD2, IFIT2, STAT1p (tyr701), STAT1, ASCC3, and cGAS in parental MRC5VA and two different CGAS knockout cell lines (KO-7 and -12) after ASCC3 knockdown. Asterisks denote non-specific bands.

(F) As in (E), but in U2OS cells.

(G) qRT-PCR analysis of relative ISG expression in U2OS cells transfected with the indicated siRNAs. Error bars represent SD of three technical replicates and are representative of three biological replicates.

(H) As in (G), but also using U2OS CGAS KO-16 cells.

See also Figures S1–S4.