Figure 8.

Immunodepletion of IFT proteins from the membrane plus matrix fraction with α-IFT72 ₁ antibody precipitates a substantial fraction of IFT motors and several axonemal proteins. Ovalbumin was added to the M+M as a negative control at 100 to 500 ng/μl before immunoprecipitation with α-IFT72₁ antibody. Pellets were resuspended in a volume equal to the supernatants and equal volume loadings of pellets (Pellet), supernatants (Super), and M+M were resolved on a 10% gel. Immunoblots show that: (A) ovalbumin did not precipitate; (B) IFT polypeptides were immunodepleted from M+M (exemplified by IFT139 and IFT74/72; the rest of IFT particle proteins were similar; unpublished data); (C) immunoblots with antibodies against IFT motors FLA10 and DHC1b, and a variety flagellar proteins as listed (Table I) show these motor proteins and various flagellar proteins coprecipitated with IFT proteins. In contrast, LC2 and Tctex-1 did not precipitate.