Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2023 Mar 2;33(4):299–311. doi: 10.1038/s41422-023-00788-1

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) under exclusive licence to Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences 2023, Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

PMC Copyright notice

Fig. 1 — a Western blotting analysis of the protein levels of cGAS in subcellular fractions including nucleus, cytosol and mitochondria in HCC cell lines. Fractionation fidelity was verified by detection of lamin B in the nuclear fraction, GAPDH in the cytosolic fraction and TOM20 in the mitochondrial fraction. b Representative images of immunofluorescence staining for cGAS in Hep3B cells with Mito-GFP expression. The nucleus was stained with DAPI. Colocalization analysis of immunofluorescence images using the colocalization plugin, which calculates Pearson’s correlation coefficient. Scale bar, 20 μm. c Western blotting analysis of the protein levels of cGAS in subcellular fractions including nucleus, cytosol and mitochondria in paired adjacent noncancerous tissues and clinical HCC tissues. Fractionation fidelity was verified by detection of lamin B in the nuclear fraction, GAPDH in the cytosolic fraction and TOM20 in the mitochondrial fraction. d Representative images of immunofluorescence staining for cGAS and TOM20 in clinical HCC tissues. The nucleus was stained with DAPI. Colocalization analysis of immunofluorescence images using the colocalization plugin, which calculates Pearson’s correlation coefficient. Scale bar, 100 μm. e Proteinase K protection assay of cGAS in the mitochondria of Hep3B or PLC cells. Left: western blotting analysis of cGAS after incubation of purified mitochondria with the indicated concentration of proteinase K. Right: proteinase K protection assays were performed in the presence of the permeabilizing agent Triton X-100. Extent of digestion was determined by blotting for key intra-mitochondrial proteins (MFN1, TOM70, COX4, TFAM). OMM, outer mitochondrial membrane; IMM, inner mitochondrial membrane; MM, mitochondrial matrix. f Co-IP assay showing the protein interaction between cGAS and TOM70. Hep3B cells were infected with lentivirus carrying Flag-EV or cGAS-Flag and HA-TOM70 plasmids. Cell lysates were immunoprecipitated with an anti-Flag antibody, followed by western blotting analysis with antibodies against Flag and HA tags. g Western blotting analysis of the protein levels of cGAS in mitochondria of Hep3B cells with TOM70 knockdown. TOM20 and GAPDH served as the loading control.