Figure EV3. USP36 promotes SUMOylation of Nop58 and Nhp2.

• A
  Candidate proteins whose SUMOylation is increased by USP36. H1299 cells transfected His‐SUMO2 together with control or Flag‐USP36 plasmids were subjected to Ni²⁺‐NTA purification followed by LC‐MS‐MS analysis.
• B, C
  USP36 SUMOylates exogenous Nop58. H1299 cells transfected with His‐SUMO2 and/or USP36 with WT Nop58 (B) or SUMOylation‐defective mutant Nop58 (K457R/K497R, Nop58^2KR) (C) were subjected to Ni²⁺‐NTA PD followed by IB to detect SUMOylation of Nop58.
• D, E
  USP36 SUMOylates exogenous Nhp2. H1299 cells transfected with His‐SUMO2 and/or USP36 with WT Nhp2 (D) or with SUMOylation‐defective mutant Nhp2 (Nop58^K5R) (E) were subjected to Ni²⁺‐NTA PD followed by IB to detect SUMOylation of Nph2.
• F
  USP36 does not SUMOylate nucleolar protein NOLC1. H1299 cells transfected with His‐SUMO2, Flag‐NOLC1 with or without V5‐USP36 were subjected to Ni²⁺‐NTA PD followed by IB to detect SUMOylation of NOLC1.
• G, H
  Co‐localization of USP36 with Nop58 and Nhp2 in the nucleolus. HeLa cells were transfected with Flag‐USP36 and stained with anti‐Flag (green) and anti‐Nop58 (red) (G) or transfected with GFP‐USP36 (green) and Flag‐Nhp2 and then stained with anti‐Flag (red) (H).

Source data are available online for this figure.