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. 2022 Oct 24;23(12):e55044. doi: 10.15252/embr.202255044

Figure EV2. CCNL1 degradation requires Cul1 and is blocked when substrate‐binding deficient FBXW7R465C is co‐expressed.

Figure EV2

  1. Quantification of immunoblots in Fig 3A, mean ± SEM, students t‐test, n = 3 independent biological replicates.
  2. Quantification of immunoblots in Fig 3D, mean ± SEM, students t‐test, n = 3 independent biological replicates.
  3. Immunoblot analysis of HEK293T cells expressing gRNAs against AAVS1 or FBXW7.
  4. TIDE analysis of HEK293T cells expressing gRNAs against AAVS1 or FBXW7.
  5. Immunoblot of immunoprecipitation of FLAG‐CCNL1 overexpressed in HPAF‐II cells, detecting endogenous Cul1 and Cul4A. Representative image of three independent replicates.
  6. Immunoblot of lysates following cycloheximide treatment of HEK293T cells expressing HA‐CCNL1 and FLAG‐FBXW7 or FLAG‐FBXW7R465C. Representative blot of three independent replicates.
  7. Quantification of cycloheximide chase in (F), mean ± SEM of three independent replicates, t‐test at T6.