The cGAS-STING-IFN pathway contributes to IL-1β secretion in response to UV-inactivated HCMV. (A) Immunoblot assay for cGAS, STING, IFNAR1, IRF3/IRF7, and GAPDH in THP-1 cells transduced with Cas9 and gRNA targeting HUMCYC pseudogene or in indicated CRISPR-generated knockout cells. WT, wild type. (B, C) Pro-IL-1β (B) and AIM2 (C) mRNA levels in THP-1 cells transduced with Cas9 and gRNA targeting the HUMCYCPS3 (ΔHUMCYC) pseudogene or indicated cellular gene(s) following treatment with UV-inactivated HCMV-TB40/E (MOI of 3) for 18 h. Transcript fold changes are expressed as relative to the levels in untreated cells (PMA−). (D) IL-1β secretion in THP-1 cells lacking indicated protein(s) following exposure to UV-inactivated HCMV-TB40/E (MOI of 3) for 18 h. Values are averages ± SEM from three biological replicates. One-way ANOVA with Bonferroni correction was performed. *, P < 0.05; **, P < 0.01; ***, P < 0.001.