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. 2023 Mar 6;24(4):e56374. doi: 10.15252/embr.202256374

Figure EV3. USP50 interacts with ACE2 and regulates ACE2 protein levels via its deubiquitinase activity.

Figure EV3

  1. IP‐IB analysis of the interaction between endogenous USP50 and ACE2 in HEK293T cells. Data are representative of three biological replicates.
  2. RT–qPCR analysis of SARS‐CoV‐2 GFP/ΔN RNA levels in 2fTGH cells transfected with control shRNAs (shCtrl) or shRNAs against USP50 (shUSP50) and then infected with SARS‐CoV‐2 GFP/ΔN (MOI = 0.1) for 2 h. Data are shown as mean and s.d. of three biological replicates (n = 3). **P < 0.01 (two‐tailed unpaired Student's t‐test).
  3. Western blot analysis of ACE2 in HEK293T cells transfected with Flag‐HA‐tagged USP50 (FH‐USP50) wild‐type (WT) or its deubiquitinase inactive mutant (C53S). Data are representative of three biological replicates.
  4. IP‐IB analysis of ubiquitination of Myc‐ACE2 in HEK293T cells cotransfected with Myc‐ACE2 and increasing amounts of Flag‐USP50. Data are representative of three biological replicates.

Source data are available online for this figure.