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. 2019 Aug 23;38(20):e101345. doi: 10.15252/embj.2018101345

Figure EV2. SCRN1 does not exhibit proteolytic activity and its C‐terminal is recruited to VAP MSP domain.

Figure EV2

  1. Schematic illustration of autolytic protease activation of C69 family members.
  2. Sequence alignment of predicted proteolytic sites of SCRN family members according to the MEROPS database.
  3. 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.
  4. 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.
  5. 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.
  6. 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).
  7. COS7 cells co‐expressing GFP‐SCRN1‐C with HA‐VAPA or HA‐VAPB. Scale bars: 10 μm (full size) and 5 μm (zoom).
  8. 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).
  9. 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.