Figure 4 .

Characterization of the MCPH1–CDH1 interaction. A) Schematic diagram of MCPH1 fragments to be used in CDH1 pull-down studies. The sites of potential D-Box and KEN-Box degron sequences are indicated. B) GFP or GFP–MCPH1 variants were transfected into HEK293 cells for 24 h before purification and incubation with FLAG–CDH1 cell extracts (from transfected HEK293 cells). The ability of CDH1 to bind MCPH1-L was investigated by using Western blotting. C) The KEN-Box sequence was mutated to “AAA” in GFP–MCPH1-L and GFP–MCPH1-S before performing the CDH1 pull-down experiment as described in B. D) Schematic diagram of CDH1 fragments to be used in pull-down studies with GFP–MCPH1-L. E) FLAG–CDH1 (or CDH1 fragment) pull-down was performed as in B.