Fig. 3
Negatively-charged residues located in the LCD are necessary for enhancing RPSAN liquefaction and protein production. A Schematic diagram of human RPSAN+IDR1 and mutant derivatives. B-D HEK-293 T cells overexpressing EGFP-RPSAWT and its mutant derivatives for 24 h. B The intracellular proteins were analyzed by immunofluorescence. Representative confocal images are shown (scale bar = 20 μm) and magnified (scale bar = 10 μm). C The mean fluorescence intensity was analyzed by flow cytometry. Quantification was performed by FlowJo as mean ± SD (n = 3 independent experiments). NS for not significant, *** for P < 0.001; one-way ANOVA with Tukey’s test. D The protein production level was detected by western blotting using the antibody against GFP