Extended Data Fig. 2. Condensate expression relocalizes tagged native clients and regulates cell growth.
a, Representative images of tagged natively expressed Cdc24 show its cortical localization. b, Representative images of tagged natively expressed Cdc5 show its punctate localization to spindle pole body. c, Images of the same Cdc24-mScarlet-TsCC(B) cell before and after induced expression of TsCC(A)-(RGG)3 scaffold for 6 hr, show loss of cortical Cdc24 signal and partitioning to synthetic condensate. d, Client recruitment to condensates specifically depends on CC tag interaction; Cdc24 does not interact with (RGG)3 condensates that lack the interaction tag. e, Left, scheme: cortical Cdc24 is relocalized from cortex to synthetic condensates after induction of scaffold. Right, Average cortical Cdc24-mScarlet-TsCC(B) signal before and after TsCC(A)- (RGG)3 scaffold expression (6h). n = 20 cells before and after hours of galactose induction. Significance calculated by unpaired, two-tailed, t-test (****, p ≤ 0.0001). f, Kinetics of loss of cortical Cdc24-mCherry-TsCC(B) signal (red) concomitant with cellular accumulation of expressed TsCC(A)-(RGG)3 scaffold (green) upon induction with galactose for 20 cells over 6 hours. Shaded area, s.d. g, Cell proliferation: measurements of cell density (OD600) over time for indicated Cdc24 strains in liquid media containing galactose. h, Growth assay for Cdc24 strains: five-fold serial dilution of indicated strains grown on solid- media containing glucose or galactose. i, Average cell area of mother cells only increases upon TsCC(A)-(RGG)3 expression in Cdc24-mScarlet-TsCC(B) cells, consistent with cell cycle arrest in G1. j, Growth assay for Cdc5 strains: five-fold serial dilution of indicated strains grown on solid-media containing glucose or galactose. In all cases, growth defect depends on presence of tagged client and expression of scaffold to form condensates. Phenotype is not observed with only native client tagging or only scaffold expression.