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. 2018 Jul 4;37(16):e98701. doi: 10.15252/embj.201798701

Figure 4. Cezanne regulates APC/C substrates degradation.

Figure 4

  1. Degradation assay of endogenous Aurora A using a G1 HeLaS3 cell extract supplemented with ATP, ubiquitin, UBE2C, and Cezanne. Aliquots were collected at the indicated time points and analyzed by immunoblot.
  2. Degradation assay was performed as in (A) except that in vitro‐translated Cyclin B was used as a substrate. Aliquots were collected at the indicated time points and analyzed by immunoblot.
  3. U2OS cells were transfected with the control firefly (FF), Cezanne, and Cezanne with UBE2S siRNAs for 48 h and analyzed by immunoblot with the indicated antibodies.
  4. U2OS cells were analyzed as in (C).
  5. U2OS cells were transfected with the indicated siRNAs, and after 48 h of knock down, an empty vector or an siRNA‐resistant Cezanne vector was introduced by transfection for an additional 24 h. Cells were analyzed by immunoblot.