Extended Data Fig. 2: LEM2 wets the surface of microtubules with a liquid-like coat.
From: LEM2 phase separation promotes ESCRT-mediated nuclear envelope reformation
a, Additional example of STED imaging of endogenous LEM2 localization during late anaphase. Scale bars, 150 nm. Representative of data from four cells. b, Fluorescence imaging of indicated combinations of LEM2FL–Alexa488 (magenta), tubulin–Alexa647 (green), and lipids labelled with PE-rhodamine (cyan). Scale bar, 2 μm. Representative of two technical replicates. c, Light scattering at 340 nm (turbidity) of microtubule bundling by indicated LEM2 constructs. Half maximal concentration of LEM2NTD is 1.303 ± 0.1 μM. Mean ± s.e.m. for n = 3 independent samples. d, Negative stain electron microscopy of microtubules alone or microtubules with indicated concentrations of LEM2NTD, corresponding to the concentrations measured by light scattering. Images representative of at least six; scale bars, 25 nm. e, Fluorescence microscopy of LEM2NTD–Alexa488 in combination with tubulin–Alexa647, LEM2WH–Alexa555, and GTP/MgCl2, as indicated. Scale bar, 10 μm. Images are representative of at least three. f, Example images for FRAP of LEM2FL- and LEM2NTD-coated microtubule bundles, representative of five independent samples (LEM2FL) or 17 independent samples (LEM2NTD). Scale bar, 2 μm. g, Top, kymograph across bleached region. Bottom, FRAP of LEM2NTD-coated microtubule bundle. Images show fluorescent LEM2NTD channel. Scale bar, 2 μm. Representative of eight technical replicates.
