Supplementary Figure 9: Full-length western blot images.

Samples were run on a 4–15% TGX SDS-gradient gel (Bio-Rad Laboratories). Molecular weight (M, in kDa): BLUEstain 2 Protein ladder, 5–245 kDA (Goldbio). Images were taken with the infrared imaging system from Odyseey (Li-cor). This allows staining for two proteins at the same time on the same plot. Light-blue bands indicate oversaturation. a. for Fig. 2e, left and right panel show the same plot, but with adjustment of brightness and contrast; b. for Fig. 2d, the second and the fourth row are adjustment in brightness and contrast from the plots in the first and the third row, respectively; c. and d. for Supplementary Fig. 7. For Syt1, we always observed in varying intensity a band at ∼75 kDa. This band is likely to be unstripped Nsf. We tried to optimize the stripping procedure, however we could never remove the strong signal band for Nsf completely. For consistency, we decided to stain always Syt1 after Nsf.