Figure S1.
PI motif of Bin1 is required for membrane binding and tubulation, but not Dyn2 fission activity. (A) Quantification of the tubulated liposome diameter in Fig. 1 H (n ≥ 11 liposome tubules). (B) Quantification of the periodicity of Dyn2 or Dyn2-Bin1 spirals assembled on liposomes in Fig. 1 H (n ≥ 11 liposome tubules). (C) Domain structure of Bin1 variants used in this study. (D) Effect of PI motif from Bin1 on regulating Dyn2 fission activity. 0.5 µM purified Dyn2 was incubated by itself or together with 0.5µM Bin1 variants with or without the PI motif in the presence of GTP and SUPER template. The data was shown as fold change relative to the Dyn2 fission activity alone (n = 3 experimental replicates). (E) Effect of PI motif on membrane tubulation ability of Bin1 in cellulo. GFP-tagged Bin1 containing PI motif (Bin1-GFP) or without PI motif (Bin1∆PI-GFP) were over-expressed in C2C12 myoblasts through transfection and viewed under confocal microscopy. Scale bar, 10 µm, scale bar for inset magnification, 5 µm. Bar graphs are shown as average ± SD, box plots are shown as median ± min and max value. Data are analyzed with Student’s t test (two-tailed, unpaired; B) or one-way ANOVA (A and D). *P < 0.05; ***P < 0.001.