FIG 2.

cGAS is required for maximal STING activation and ISG responses to VacV lacking F17 in human MRC5 lung fibroblasts. (A) Control or two independent lung fibroblast MRC5 cGAS KO cell lines were mock infected (M) or infected with WT or iF17 virus at an MOI of 5 for 30 h. Whole-cell lysates were subjected to Western blotting. int, intermediate; U>, unprocessed A10; P>, processed A10. (B) Differential abundances of DNA sensors in human cells. Unmodified THP1 cells, commercial THP1 Dual reporter cells used in the experiment whose results are shown in Fig. 1, MRC5 cells, and primary NHDF cells were mock infected (M) or infected with WT or iF17 virus at an MOI of 5 for 40 h (THP1 cells), 48 h (THP1 reporter cells), or 30 h (MRC5 and NHDF cells). Whole-cell lysates were subjected to Western blotting with antibodies to the indicated proteins. Note that cGAS is expressed at far lower levels and degradation of a subpopulation of cGAS is more readily detectable in fibroblasts than in THP1 cells. All data are derived from and representative of at least 3 biological replicates.