cGAS and STING are recruited to micronuclei.

(A) Mouse fibroblasts were transfected with plasmids encoding mCherry or mCherry-cGAS (magenta), treated with 4-OHT (iKO), fixed at 1-2d, and stained with DAPI (DNA; blue). Arrowheads point to a cGAS-positive micronucleus. Scale bars, 20μm. (B) Cells were treated with DMSO (Flox) or 4-OHT (iKO) for 3-4d and immunoblotted with antibodies to cGAS, STING, ArpC2, tubulin, actin, and GAPDH. (C) cGAS and STING band intensities were normalized to tubulin, actin, and GAPDH band intensities. Each bar represents the mean intensity ±SD from n = 3 or 6 experiments. (D) Flox and iKO cells were fixed at 3d and stained with cGAS (magenta) or STING (green) antibodies and DAPI. Arrowheads point to cGAS- or STING-positive micronuclei. 38.7% of micronuclei in iKO cells were cGAS-positive (75 cells evaluated from 2 experiments). Insets show STING localization around an iKO micronucleus. (E) iKO cells were fixed at 6d and stained with antibodies to STING (green) and either GRP94 or GM130 (yellow).

Read more here: Source link