Extended Data Fig. 7: Dissolution of membraneless organelles is dependent on DYRK3 localization and its kinase activity. | Nature

Extended Data Fig. 7: Dissolution of membraneless organelles is dependent on DYRK3 localization and its kinase activity.

From: Kinase-controlled phase transition of membraneless organelles in mitosis

Extended Data Fig. 7

a, Images show cells overexpressing wild-type EGFP–DYRK3(WT) and kinase-dead EGFP–DYRK3(K218M). These images are the same as Fig. 3a showing both channels. b, Dissolution of splicing speckles upon overexpression of EGFP–DYRK3(WT) in interphase cells is reversed by GSK-626616 treatment (1 μM, 2 h). c, Top, schematic of the EGFP-tagged nuclear localization signal (NLS) mutant of DYRK3 (EGFP–NLSmut-DYRK3). Arginine and lysine residues in the NLS are mutated to alanine. Bottom, overexpressed EGFP–NLSmut-DYRK3 localizes to the cytoplasm and does not dissolve splicing speckles. d, Left, dissolution of splicing speckles upon overexpression of EGFP–NLS(SV40)-DYRK3(WT) in interphase cells. Middle, dissolution of splicing speckles upon overexpression of EGFP–NLS(SV40)-DYRK3(WT) in interphase cells is reversed upon GSK-626616 treatment (1 μM, 2 h). Right, condensed splicing speckles upon overexpression of EGFP–NLS(SV40)-DYRK3(K218M) in interphase cells. e, Images show cells overexpressing EGFP–DYRK3(WT) and EGFP–DYRK3(K218M). These images are the same as Fig. 3c showing both channels. f, Dissolution of pericentriolar satellites upon overexpression of EGFP–DYRK3(WT) in interphase cells is reversed upon GSK-626616 treatment (1 μM, 2 h). g, Images show cells overexpressing EGFP–DYRK3(WT) and EGFP–DYRK3(K218M). These images are the same as Fig. 3e showing both channels. h, Dissolution of stress granules upon overexpression of EGFP–DYRK3(WT) in interphase cells is reversed upon GSK-626616 treatment (1 μM, 2 h). i, The percentage of low complexity regions (LCR) occupying each full-length protein was computed for all DYRK3 interactors (Fig. 1a) and all the known splicing-speckle components (GO:0016607). SRRM1 shown in blue is among the proteins with the highest proportion of low complexity regions. j, Dissolution of EGFP–PCM1(1–1468) cytosolic granules upon overexpression of RFP–DYRK3(WT), and not RFP–DYRK3(K218M). k, Time-lapse images show RFP–DYRK3(WT) driven dissolution of cytosolic granules formed upon EGFP–PCM1(1–1468) overexpression. l, Dissolution of mCherry–SRRM1 nuclear granules upon overexpression of EGFP–DYRK3(WT), and not EGFP–DYRK3(K218M). m, Nucleoli are unaffected upon overexpression of EGFP–DYRK3(WT) or EGFP–DYRK3(K218M) in interphase cells. Images are representative of at least three independent experiments. Scale bars, 10 μm.