Extended Data Fig. 1: cGAS translocates to the nucleus in non-apoptotic cells with intact nuclear membranes. | Nature

Extended Data Fig. 1: cGAS translocates to the nucleus in non-apoptotic cells with intact nuclear membranes.

From: Nuclear cGAS suppresses DNA repair and promotes tumorigenesis

Extended Data Fig. 1

ae, Immunofluorescence results depicting localization of endogenous cGAS (anti-cGAS) in HCA2-TERT cells, an TERT-immortalized human fibroblast line. Cells were untreated (NT) (a) or exposed to etoposide (100 μg ml−1) (b), camptothecin (1 μM) (c) or H2O2 (10 mM) (d) for the indicated times. Data are representative of n = 3 independent experiments. Quantitative data are shown in e. At least 100 cells were counted in each experiment. Data are mean ± s.e.m. of n = 3 independent experiments. fi, Results of immunofluorescence analysis showing localization of overexpressed HA–cGAS in human HCA2-TERT fibroblasts that were untreated (NT) (f) or exposed to camptothecin (1 μM) (g) or H2O2 (10 mM) (h) for the indicated times. Data are representative of n = 3 independent experiments. Quantitative data are shown in i. At least 100 transfected cells were counted in each experiment. Data are mean ± s.e.m. of n = 3 independent experiments. jn, Representative immunofluorescence analysis showing the localization of HA–cGAS (anti-HA, green) in transfected primary human skin fibroblasts from one healthy donor that were untreated (NT) (j) or exposed to etoposide (100 μg ml−1) (k), camptothecin (1 μM) (l) or H2O2 (10 mM) (m) for the indicated times. Data represent n = 2 independent experiments. The quantification of nuclear translocation of cGAS in primary skin fibroblasts from two donors in response to indicated DNA-damaging agents are shown in n. At least 100 transfected cells were counted in each experiment. o, Representative FACS results depicting apoptosis of human fibroblast HCA2-TERT cells treated with etoposide (100 μg ml−1), camptothecin (1 μM), H2O2 (10 mM) or carbonyl cyanide 3-chlorophenylhydrazone (CCCP) (10 μM) for the indicated times. Data represent n = 3 independent experiments. Scale bar, 5 μm.