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. 2024 Dec 2;134(23):e181044. doi: 10.1172/JCI181044

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© 2024 Zhang et al.

This work is licensed under the Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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(A and C) Cytoplasmic and nuclear protein fractions were extracted for immunoblot analysis to determine the subcellular localization of cGAS in HK1 cells with the indicated cell lines and stimulation. (B and D) Subcellular distribution (cytoplasm and nucleus) of cGAS was determined with immunofluorescence staining of HK1 cells with the indicated cell lines and stimulation (scale bars: 10 μm). The percentages of cells (n = 200) in the nucleus, cytoplasm, or both the cytoplasm and nucleus were calculated (E) Confocal microscopy images of cGAS and HO-1 in HK1 cells before and after RT (scale bars: 10 μm). (F) The cytoplasmic and nuclear protein fractions of HK1 cells at the indicated RT time points were extracted for coimmunoprecipitation. (G and H) The interaction of HA-tagged full-length cGAS (aa 1–522), N-terminus of cGAS (aa 1–160), C-terminus of cGAS (aa 161–522), and Flag-tagged HO-1ΔTMS in HEK293T cells was analyzed by immunoprecipitation. (I) HMOX1-KO HK1 cells were stably transfected with cleaved HO-1 (HO-1ΔTMS). The interaction of endogenous cGAS and CRM1 in HK1 cells was analyzed by immunoprecipitation. All representative data from 1 experiment are shown (n = 3 biologically independent experiments).