Extended Data Fig. 2: LATS2 as scaffold protein mediates its condensations by PRM.

a, The puncta quantitation of AD293 cells transfected by mEGFP, mEGFP-LATS2, Flag-LATS2, and HA-LATS2. n = 5 cells, thee histogram shows the puncta number of individual AD293 cells; the scatter plot shows the puncta size of individual AD293 cells with indicated plasmids transfection. b, Live imaging assay and typical images of AD293 cells with transfection of mEGFP-LATS2. The red rectangles labeled the fusion events, also see Supplementary Video 3. c, The puncta quantitation of AD293 cell with transfection of mEGFP-LATS2 treated by DMSO, 3% 1,6 Hex, and 3% 2,5 Hex respectively. The percentage of cells with cytoplasmic puncta were acquired from three random views. The puncta number were calculated in 5 cells. d, The puncta quantitation of AD293 cells expressing indicated LATS2 deletions (green) and stained for nuclei. The histograms showed the mean percentage of cell with puncta from three random views and mean puncta in 5 cells; the scatter plot showed the puncta size of indicated mEGFP-LATS2 deletions in 5 cells. e, The puncta quantitation of AD293 cells expressing indicated LATS2 mutations (green) and stained for nuclei. The histograms showed the mean percentage of cell with puncta from three random views and mean puncta number in 5 cells. f, FRAP assay of AD293 cells expressing LATS2-PRM-mCherry-Cry2. The representative images of pre-bleach, bleach, and post-bleach were shown and the red and yellow rectangles marked the bleach and control puncta. The plot is the quantitation of dynamic intensity of bleached and control puncta. The experiments were performed three independent times. Also see Supplementary Video 8. g, Live imaging traces the LATS2-PRM-mCherry-Cry2 puncta after 488 nm laser illumination. The representative images were shown and the green rectangle marked the fusion puncta. Also see Supplementary Video 8. h, FRAP assay of LATS2-PRM -Cy3. The representative images of pre-bleach, bleach, and post-bleach were shown and the red and yellow rectangles marked the bleach and control puncta. The plot is the quantitation of dynamic intensity of bleached and control puncta. The experiments were performed three independent times; also see Supplementary Video 10. Data are presented as mean ± SEM. Significant differences are labelled; One-way ANOVA was used for c statistical analysis. ***P < 0.001, ****P < 0.0001, ****P < 0.0001. Scale bars, 5 µm or 10 µm as indicated.