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. Author manuscript; available in PMC: 2022 Dec 24.
Published in final edited form as: Biochemistry. 2022 Jun 23;61(14):1495–1507. doi: 10.1021/acs.biochem.2c00118

Figure 1. ER-derived microsomal structures and the association of FL SUN proteins with membrane fragments in HeLa cell lysates used for CFE.

Figure 1.

A) Illustration of a SUN2 homo-trimer embedded in the INM that is interacting with KASH proteins embedded in the ONM. B) Schematic illustrating the use of an Airfuge to spin down ER-derived microsomal membranes in bulk CFE reactions. C) Representative in-gel fluorescence images of EGFP and the indicated EGFP-tagged proteins in supernatant and pellet fractions post ultracentrifugation. A self-inserting mechanosensitive membrane channel MscL was also expressed and served as a positive control. D) Representative confocal fluorescence images of ER tracker-labeled HeLa lysate before and after spinning down microsomal membranes. Scale bar: 1 μm. E) Cryo-EM image of HeLa CFE system expressing EGFP-SUN1FL showing vesicular structures. Small structures are visible in the background indicating ribosomes (~ 30 nm). Scale bar: 100 nm.