Extended Data Fig. 8: MeCP2 Mini forms droplets in vitro and partitions into heterochromatin condensates in mouse ES cells.
From: MeCP2 links heterochromatin condensates and neurodevelopmental disease
a, Schematic of MeCP2 protein with a minimal MeCP2 protein (Mini)22 that retains the MBD and NID displayed below. b, Droplet experiments examining ability of Mini MeCP2 to form droplets. Wild-type MeCP2–GFP and Mini at 4 μM were added to droplet formation buffers with 125 mM NaCl and 10% PEG-8000. c, Droplet areas for experiments in b. Fields per condition n = 12. d, MeCP2–GFP condensed fraction for experiments in b. Fields per condition n = 12. e, Droplet experiments examining ability of wild-type MeCP2 and Mini to form droplets with HP1α and DNA. Wild-type MeCP2–GFP or Mini at 7.5 μM was mixed with 7.5 μM HP1α–mCherry and 160 nM DNA in droplet formation buffers with 150 mM NaCl and 10% PEG-8000. f, HP1α–mCherry partition ratios in wild-type MeCP2–GFP and Mini droplets for experiments in e. Fields per condition n = 15. g, DNA-Cy5 partition ratios in wild-type MeCP2–GFP and Mini droplets for experiments in e. Fields per condition n = 15. h, Droplet experiments examining ability of MeCP2 wild-type and Mini condensates to enrich TBLR1-CTD. Wild-type MeCP2–GFP or Mini at 4 μM was mixed with TBLR1-CTD-mCherry at 10 μM in droplet formation buffers with 125 mM NaCl and 10% PEG-8000. i, TBLR1-CTD-mCherry partition ratios in wild-type MeCP2–GFP and Mini droplets for experiments in h. Fields per condition n = 12. j, Live-cell microscopy of endogenous-tagged wild-type MeCP2–GFP and Mini proteins with Hoechst staining in mouse ES cells. k, Partition ratios of MeCP2–GFP proteins at heterochromatin condensates for experiments in j. n = 10 cells per condition. P = 0.1161, t = 1.6509, df = 18, two-tailed Student’s t-test. All data are mean ± s.d.
