Fig. 1: GEM2 labeling of mitochondrial surface-displayed EGFP in human cells.
From: Genetically encoded multimeric tags for subcellular protein localization in cryo-EM
a, Schematic of the labeling system. b, Time course of GEM2 (fluorescently labeled with TMR, magenta) recruitment to Mito-EGFP (green) upon rapalog treatment by fluorescence microscopy in HeLa cells. GEM2 and adaptor protein were expressed from the AAVS1 locus (stable knock-in) with 24–48 h doxycycline induction before rapalog treatment. c, Quantification of b, showing the fraction of GEMs overlapping with Mito-EGFP per cell. Lines indicate mean, n (left to right) = 40, 40, 41, 40 cells, two experiments. ***P < 0.0001, Kruskall–Wallis test followed by Dunn’s test, compared to 0 min. d, Tomographic slice showing a GEM2-labeled mitochondrion after 30 min rapalog treatment. Arrowheads in the inset indicate GEM2 particles.
