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. 2023 Nov 21;24(12):e56920. doi: 10.15252/embr.202356920

Figure EV1. The UFL1‐DDRGK1 complex.

Figure EV1

  • A
    Model confidence of the UFL1‐DDRGK1 AlphaFold2 complex prediction. Left: Predicted local distance difference test (pLDDT); Right: predicted Align Error (pAE) plots of model 1, and models 2–5. For visualization purposes, only rank_1 was used in Fig 1 (accompanies Fig 1A).
  • B
    Model of the DDRGK1‐UFL1 complex, colored according to pLDDT, highlighting the lower confidence in the structure of the N‐terminal helix.
  • C
    SDS–PAGE showing the purity of the indicated fusion proteins.
  • D–H
    Loading controls of in vitro ufmylation assays (accompanies Fig 1E–H). (D, E) fusion constructs, (F, G) ternary complex. (H) Presence of UFL1/UFL1ΔN in the membrane: Since we hardly see UFL1/UFL1ΔN in the Ponceau staining (F), we performed Western blot analysis with anti‐Myc. All reactions were loaded on bis‐Tris‐PAGE (except the reaction of Fig 1G, which was loaded on 8–16% Tris gel).