Feline Lindhout

VAP–SCRN1 interaction regulates dynamic endoplasmic reticulum remodeling and presynaptic function 143 4 Supplementary Figure 2. SCRN1 does not exhibit proteolytic activity and its C-terminal is recruited to VAP MSP domain A. Schematic illustration of autolytic protease activation of C69 family members. B. Sequence alignment of predicted proteolytic sites of SCRN family members according to the MEROPS database. C. Western blot of lysates from HEK293T cells expressing BioGFP-SCRN1-WT, BioGFP-SCRN1- C9A, BioGFP-SCRN2-WT, BioGFP-SCRN2-C12A, BioGFP-SCRN3-WT, or BioGFP-SCRN3-C6A. Arrowheads represent (1) full-length BioGFP-SCRN proteins and (2) N-terminal cleaved BioGFP- SCRN2 and BioGFP-SCRN3. D. Pull-down assay of HEK293T cells co-expressing Myc-VAPA with BioGFP, BioGFP-SCRN1-WT, BioGFP-SCRN1-C9A, BioGFP-SCRN1-N, or BioGFP-SCRN1-C. E. Pull-down assay of HEK293T cells co-expressing Myc-VAPB with BioGFP, BioGFP-SCRN1-WT, BioGFP-SCRN1-C9A, BioGFP-SCRN1-N, or BioGFP-SCRN1-C. F. Hippocampal neurons (DIV16) co-expressing HA-VAPB with GFP-SCRN1-N or GFP-SCRN1-C. Scale bars: 10 μm (full size) and 2 μm (zoom). G. COS7 cells co-expressing GFP-SCRN1-C with HA-VAPA or HA-VAPB. Scale bars: 10 μm (full size) and 5 μm (zoom). H. COS7 cells co-expressing HA-SCRN1 with GFP-VAPB-TM, GFP-VAPB-MSP-CC, or GFP-VAPB- MSP. Scale bars: 10 μm (full size) and 5 μm (zoom). I. COS7 cells co-expressing GFP-SCRN1-N with HA-VAPA or HA-VAPB. Scale bars: 10 μm (full size) and 5 μm (zoom). Source data are available online for this Figure.