Fig. 3: Heterotypic PLDs interact and enrich within homotypic PLD condensates.

a A Schematic of the co-partitioning assay based on confocal fluorescence microscopy. 50 µM concentration of the scaffold^PLD was used to form condensates and ~1 µM of the AlexaFluor488 labeled client^PLD was utilized to determine the partition coefficient (\(k\)). Created with BioRender.com. b Partitioning of AlexaFluor488 labeled client PLDs (ARID1A^PLD, ARID1B^PLD, SS18^PLD, BRG1^PLD, RNA Polymerase II CTD³⁰, and FUS^PLD) within condensates of SS18^PLD. Enrichment (partition coefficient) is calculated as shown in (a) and displayed as a box-and-whisker plot. (ARID1A^PLD n = 235 condensates, ARID1B^PLD n = 222 condensates, SS18^PLD n = 268 condensates, BRG1^PLD n = 326 condensates, RNA Polymerase II CTD³⁰ n = 450 condensates, and FUS^PLD n = 385 condensates). The scale bar is 10 μm. c The average partition coefficient (\(k\)) is tabulated along with the standard deviation from the mean. d HEK293T cells co-expressing GFP-SS18^PLD and either one of the mCherry-tagged PLDs (ARID1A^PLD, ARID1B^PLD, FUS^PLD and BRG1^PLD) or mCherry alone. The degree of colocalization is displayed as intensity profiles for condensates shown in the inset images. Green represents the intensity profile of GFP-SS18^PLD and red represents the profile for mCherry-tagged PLDs. The enrichment coefficients are reported in Fig. S9. The scale bar is 10 μm.