Fig. 1
IDR of RPSA is necessary for liquid-like condensation in HEK-293 T cells. A The C-terminal (207 AA-295 AA) of human RPSA is intrinsically unstructured (https://iupred2a.elte.hu/). B-D HEK-293 T cells overexpressing human EGFP-RPSAWT, EGFP-RPSAN1 AA−206 AA and EGFP-IDR207 AA−295 AA for 24 h. B FRAP analyses of RPSA condensates. A typical condensate is shown (scale bar = 20 μm) and magnified (scale bar = 10 μm). C FRAP analyses of RPSAN and IDR condensates. FRAP images of condensates are shown (scale bar = 20 μm) and magnified (scale bar = 5 μm). Quantification of FRAP on condensates (n = 5 condensates) is shown (mean ± SD). (B and C) "CI" denotes a continuous imaging combination. "Bleached" denotes the extent of fluorescence quenching, and subsequent continuous shooting is used to measure the extent of fluorescence recovery. D Representative confocal images are shown (scale bar = 20 μm) and magnified (scale bar = 10 μm). The cell membrane was not permeabilized while labeling GFP proteins with 594-conjugated antibodies. Areas of 594-GFP on the cell surface were used for quantitative analysis by Image J (RPSAN group: n = 30 cells, IDR group: n = 40 cells, mean ± median). ** for P < 0.01; Mann–Whitney test