Fig. 6: CEP112 undergoes liquid-liquid phase separation (LLPS) in vitro and in vivo.

a, b Fluorescence microscopy reveals that GFP-CEP112 forms condensates within HEK293T cells (a). 3D visualization confirms the spherical morphology of these intracellular condensates (b) (Scale bar: 5 μm). c Time-lapse fluorescence microscopy demonstrates that GFP-CEP112 condensates undergo rapid fusion and fission events in HEK293T cells. Magnified areas are indicated by purple dotted boxes. d Fluorescence recovery after photobleaching (FRAP) assay of GFP-CEP112 droplets in HEK293T cells. Representative images (left) show bleaching at the indicated time points and recovery at 37 °C (Scale bar: 5 μm). Fluorescence-intensity analysis (right) of the FRAP data over 300 s. Three independent experiments were performed. Data are presented as mean values ± SEM. Source data are provided as a Source Data file. e GFP-CEP112 undergoes LLPS in vitro in a protein and salt concentration-dependent manner. Fluorescence microscopy images show the formation of GFP-CEP112 droplets at increasing protein concentrations (top) and NaCl concentrations (left) (Scale bar: 10 μm). f FRAP assay of in vitro GFP-CEP112 droplets. Representative images (left) depict bleaching at the indicated time points and recovery at 37 °C (Scale bar: 5 μm). Fluorescence intensity analysis (right) of FRAP data over 300 s. Three independent experiments were performed. Data are presented as mean values ± SEM. Source data are provided as a Source Data file. g Confocal microscopy reveals co-localization of GFP-CEP112 (green) and mCherry-tagged hnRNPA2B1, eIF4A1, or eEF1A1 (red) within droplets in vitro (Scale bar: 10 μm). h Purified GFP-CEP112 selectively recruits its target RNAs, FSIP2-CDS and FSIP2-5’UTR, but not the control METTL5-CDS, into phase-separated droplets in vitro (Scale bar: 10 μm). Experiments were repeated three times independently with similar results; representative images are shown in a, b, e, g and h.