Panel A shows four sets of microscopy images with EGFP, mCherry-CAST, and merged channels. Each set includes EGFP alone, EGFP-Endophilin-A1, EGFP-Endophilin-A2, and EGFP-Endophilin-A3 expressed with mCherry-CAST. The images display co-condensates formation. The right side of Panel A includes line scans of fluorescence intensity and dot plots showing the ratio of average fluorescence intensity at the condensate to the cytosolic region. Panel B shows four sets of microscopy images with EGFP-Endophilin-A2, EGFP-Endophilin-A2-BAR, EGFP-Endophilin-A2-SH3, and EGFP alone expressed with mCherry-CAST. The right side of Panel B includes line scans of fluorescence intensity and dot plots showing the ratio of average fluorescence intensity at the condensate to the cytosolic region. The graphs and images analyze the interaction efficiency of different EGFP-Endophilin proteins with mCherry-CAST.
Endophilin-A co-condensates with CAST/ELKS in HEK293T cells. (A and B) EGFP-endophilin-A1, -A2, or -A3 expressed with mCherry-CAST resulted in co-condensates in the HEK cells. EGFP alone with mCherry-CAST (upper images) did not result in condensate structures. The dotted line indicates a line scan of each fluorescence protein. Right dot plots show the ratio of average fluorescence intensity at the condensate (B) to cytosolic region (A) Scale bar: 10 μm. n = 10 cells, each. P values were obtained by comparison with EGFP-C1–expressing cells (one-way ANOVA with post hoc Bonferroni’s multiple comparison test, *P < 0.05, **P < 0.01, ****P < 0.0001). (B) EGFP–endophilin-A2 and EGFP–endophilin-A2-BAR expressed with mCherry-CAST resulted in condensates. EGFP–endophilin-A2-SH3 or EGFP alone expressed with mCherry-CAST did not result in condensates. The dotted line indicates a line scan of each fluorescence protein. Right dot plots show the ratio of average fluorescence intensity at the condensate (B) to cytosolic region (A). Scale bar: 10 μm. n = 10 cells, each. P values were obtained by comparison with EGFP–endophilin-A2–expressing cells (one-way ANOVA with post hoc Bonferroni’s multiple comparison test, ***P < 0.001).