Fig 1: Liprin-α1 SLiM4 mutant significantly increased GFP-liprin-α1 liquid-liquid phase separation.A, representative live confocal images of WT HEK293 cells transfected with GFP-liprin-α1 (WT), GFP-liprin-α1 SLiM1 mutant (SLiM1), GFP-liprin-α1 SLiM4 mutant (SLiM4), or GFP-liprin-α1 SLiM1, and 4 mutant (SLiM1+4). The scale bar represents 20 μm. B, quantification of the percentage of WT HEK293 cells with droplets. Shown are mean ± SD as well as individual data points from three technical replicates. Significance was determined by one-way ANOVA with Dunnett's multiple comparison post hoc test (ns: not significant, ∗∗∗p ≤ 0.001). C, an example of time-lapse confocal images of WT HEK293 cells transfected with SLiM4. Droplets subject to fluorescence recovery after photobleaching (FRAP). One hundred twenty frames acquired with 0.5 s between each frame. Bleaching occurred at frame 2 (t = 0 s). Images show the droplet directly before and after bleaching as well as the fluorescence recovery over time. D, normalized percentage of FRAP plotted over time. N = 30 droplets/3 technical replicates. Error bars indicate mean ± SD. SLiM, short linear motif.