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. Author manuscript; available in PMC: 2020 Feb 26.
Published in final edited form as: Nat Immunol. 2019 Aug 26;20(10):1381–1392. doi: 10.1038/s41590-019-0469-z

Figure 5.

Figure 5.

Fbxl1 and Fbxl12 target the same site on Cdkn1b for K48 poly-ubiquitination. (a) Immunoprecipitation and immunoblot analysis showing K-48 but not K-63 ubiquitinylation of Cdkn1b by SCF-Fbxl12 complexes in HEK-293T cells transfected for 48 h with plasmids encoding Myc-Fbxl12, Flag-Cdkn1b and either HA-UbK48 or HA-UbK63 followed by treatment with MG132 for 8 h. (b) Immunoprecipitation and immunoblot analysis showing K-48 but not K-63 ubiquitinylation of Cdkn1b by SCF-Fbxl1 complexes in HEK-293T cells transfected for 48 h with plasmids encoding Myc-Fbxl1, Flag-Cdkn1b and either HA-UbK48 or HA-UbK63 followed by treatment with MG132 for 8 h. (c) Immunoprecipitation and immunoblot analysis showing ubiquitinylation of Cdkn1b at K165 by SCF-Fbxl1 and SCF-Fbxl12 complexes in HEK-293T cells transfected with plasmids encoding HA-Ub, Flag-Cdkn1b or Flag-Cdkn1b(K165R) and Myc-Fbxl1 or Myc-Fbxl12 for 48 h then treated with MG132 for 8 h. Results shown are representative of at least three independent experiments.