Supplementary Figure 7: Monomeric and fusion properties of mEosEM.
From: mEosEM withstands osmium staining and Epon embedding for super-resolution CLEM
a, Size-exclusion chromatography of purified mEosEM and mEos4b (10 μM). b, Presentation of OSER assay of mEosEM. Left, normal structure of CytERM-mEosEM. Right, abnormal structure of CytERM-mEosEM. The white arrow indicates the whorl structure. Scale bars, 15 μm. The OSER assay1 of mEosEM showed that 97% of the cells exhibited normal CytERM structures (without nuclear envelope thickenings, whorl structures), n = 1532 cells. c, Sedimentation velocity analytical ultracentrifugation of mEosEM and mEos4b proteins (0.05 mM). The theoretical molecular weight of monomeric of mEosEM and mEos4b with a 6 × His tag is approximately 30 kD. d, Confocal images of mEosEM-labeled structures. Lamin A (i), H2B (ii), F-actin (iii), mitochondria (iv), CytERM (v) and caveolin (vi). Scale bars, 10 μm. The experiments of a, b and d were performed three times with similar results. 1. Costantini, L.M., Fossati, M., Francolini, M. & Snapp, E.L. Assessing the Tendency of Fluorescent Proteins to Oligomerize Under Physiologic Conditions. Traffic 13, 643-649 (2012).
