Fig. 5: ZNF252P directly drives phase separation of HNRNPK and ILF3 in vitro.

A Graph plotting intrinsically disordered regions (IDRs) of HNRNPK and ILF3 by PONDR. B Representative images and quantification of formed droplets of GFP-HNRNPK/mCherry-ILF3 at the indicated salt concentration in vitro. HNRNPK protein concentration: 10 μM. ILF3 protein concentration: 4 μM. Scale bar 10 μm. Data are representatives or mean ± s.d (Left panel, 0 mM: n = 13 droplets (HNRNPK); 100 mM: n = 31 droplets (HNRNPK); 200 mM: n = 112 droplets (HNRNPK). 0 mM: n = 5 droplets (GFP-HNRNPK); 100 mM: n = 32 droplets (GFP-HNRNPK); 200 mM: n = 48 droplets (GFP-HNRNPK). Right panel, 0 mM: n = 15 droplets (ILF3); 100 mM: n = 43 droplets (ILF3); 200 mM: n = 108 droplets (ILF3). 0 mM: n = 8 droplets (mCherry-ILF3); 100 mM: n = 36 droplets (mCherry-ILF3); 200 mM: n = 68 droplets (mCherry-ILF3).Three independent experiments were performed, one-way ANOVA with Dunnett’s multiple comparisons test. C The fluorescence intensity recovery of GFP-HNRNPK and mCherry-ILF3 droplets recovered after photobleaching in vitro. Scale bar 1 μm. Data are representatives or mean ± s.d.; n = 3 independent experiments. D The fluorescent detection image of Cy3-labeled ZNF252P. Data are representatives of 3 independent experiments. E The formation and quantification of HNRNPK or ILF3 droplets in the presence of different concentrations of Cy-3 labeled ZNF252P RNA. Scale bar 10 μm. Data are representatives or mean ± s.d.; n = 3 independent experiments, one-way ANOVA with Dunnett’s multiple comparisons test. F Representative images and quantification of the effect of ZNF252P deletion on HNRNPK or ILF3 droplets formation in the droplet assay. Scale bar 10 μm. Data are representatives or mean ± s.d.; n = 3 independent experiments, one-way ANOVA with Dunnett’s multiple comparisons test. Source data are provided as a Source data file.