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. 2021 Apr 14;22(6):e50684. doi: 10.15252/embr.202050684

Figure EV1. USP36 promotes SUMOylation in cells.

Figure EV1

  • A., B
    USP36 promotes SUMOylation in cells. 293 (A) and T47D (B) cells transfected with control or Flag‐USP36 plasmid were assayed by IB for SUMOylated proteins.
  • C, D
    USP36 promotion of SUMOylation is specific. H1299 cells transfected with increasing amounts of Flag‐USP36, Flag‐JOSD3 (C), or Flag‐USP7 (D) were assayed by IB.
  • E, F
    USP36 promotes SUMOylation by SUMO1. H1299 cells (E) or HeLa and U2OS cells (F) transfected with Flag‐USP36 were assayed by IB using anti‐SUMO1 antibody.
  • G
    USP36 promotes SUMOylation by exogenous SUMO2. H1299 cells transfected with His‐SUMO2 and/or Flag‐USP36 were subjected to Ni2+‐NTA agarose beads pulldown (PD) followed by IB using anti‐SUMO2/3 antibody (top panel). WCL were also directly assayed by IB using indicated antibodies (bottom panel).
  • H
    Nucleolar localization of USP36. HeLa cells transfected with Flag‐USP36 were stained with anti‐Flag (red) and anti‐B23 (green) antibodies followed by DAPI (blue).
  • I
    USP36 promotes nucleolar SUMOylation. HeLa cells transfected with control or V5‐USP36 were fractionated to the cytoplasm (Cyto), nucleoplasm (Np), and nucleolus (No) fractions, followed by IB to detect the indicated proteins.

Source data are available online for this figure.