Figure EV5. Cep78 and VprBP are independently recruited to the centrosome and Cep78 is not an EDD‐DYRK2‐DDB1^{Vpr BP}substrate.

• A
  RPE‐1 cells transfected with NS siRNA, Cep78 siRNA, or VprBP siRNA were stained with DAPI (blue) and antibodies against centrin (green) and Cep78 or VprBP (red). Scale bar, 1 μm.
• B
  Flag‐Cep78 was expressed in HEK293 cells. Lysates were immunoprecipitated with an anti‐Flag antibody in the presence or absence of 1% SDS and Western blotted with the indicated antibodies. IN, input.
• C
  HEK293 cells expressing Flag or Flag‐Cep78, HA or HA‐Ub, and Myc or Myc‐VprBP were synchronized in mitosis with nocodazole. Lysates were immunoprecipitated with an anti‐Flag antibody in 1% SDS and Western blotted with the indicated antibodies. IN, input.
• D, E
  HA‐Ub, Flag‐katanin p60 or Flag‐TERT, and Myc or Myc‐VprBP were co‐expressed in HEK293 cells. Lysates were immunoprecipitated with an anti‐Flag antibody in 1% SDS and Western blotted with the indicated antibodies. IN, input.
• F
  HEK293 cells transfected with NS siRNA or VprBP siRNA were synchronized in mitosis. Lysates were Western blotted with the indicated antibodies. α‐tubulin was used as a loading control.
• G
  HEK293 cells expressing Flag, Flag‐Cep78 wild type (1–722), or mutant (1‐722D290A) were synchronized in mitosis. Lysates were immunoprecipitated with an anti‐Flag antibody in 1% SDS and Western blotted with the indicated antibodies. IN, input.

Source data are available online for this figure.