Fig. 4.

Cep44 specifically binds to and stabilizes rootletin, and its localization is independent of known linker proteins. (A) U2OS cells transfected with non-specific (NS), C-Nap1, LRRC45, rootletin, Cep215 or Cep68 siRNAs were stained with the indicated antibodies. Scale bar: 10 µm. (B) U2OS cells were transfected with NS or Cep44 siRNAs (oligo 1 or 2). Lysates were western blotted with the indicated antibodies. α-tubulin was used as loading control. (C) U2OS cells were transfected with NS, C-Nap1, LRRC45, rootletin, Cep215 or Cep68 siRNAs. Lysates were western blotted with the indicated antibodies. α-tubulin was used as loading control. (D) Western blotting of endogenous Cep44, rootletin and C-Nap1 after immunoprecipitation from U2OS lysates with control, anti-Cep44 or anti-rootletin antibodies. IN, input (5 %); IP, immunoprecipitation. (E) U2OS cells were transfected with NS or Cep44 siRNAs and treated with vehicle, MG132 or chloroquine. Lysates were western blotted with the indicated antibodies. α-tubulin was used as loading control.