Fig. 4

ENO stimulation promotes RPSA liquid-like condensation. A HCMEC/D3 cells were treated with or without 1,6-Hex. Immunofluorescence shows the degree of aggregation of RPSA on the cell surface (white arrows). Representative confocal images are shown (scale bar = 10 μm). B and C HEK-293 T cells overexpressing EGFP-RPSA.^WT for 12 h, and then ENO (20 μg/mL) protein was added to the transfected cells. After 12 h, cells were treated with or without 1,6-Hex. B Immunofluorescence analysis of the size of RPSA condensates (white arrows show condensates). Representative confocal images are shown (scale bar = 20 μm) and magnified (scale bar = 2 μm). The area of condensates was used for quantitative analysis by Image J (Control group: n = 18 condensates, ENO group: n = 25 condensates, ENO + 1,6-Hex group: n = 18 condensates, mean ± median). ** for P < 0.01, *** for P < 0.001; one-way ANOVA with Kruskal–Wallis test. C FRAP analyzes indicated the recovery ability of condensates. FRAP images are shown (scale bar = 20 μm) and magnified (scale bar = 5 μm). In Fig. 1, the symbols for "CI" and "Bleached" were depicted. Quantification of FRAP on condensates (n = 10 condensates) is shown (mean ± SD)