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Figure S2. NEDD1 copurifies with the γTuRC. HeLa extract fractionated by gel filtration using a Superose 6 column in IP buffer containing 10% glycerol. Proteins were precipitated from the fractions with methanol; separated by gel electrophoresis; blotted; and probed with antibodies against GCP2, GCP4, NEDD1, and γ-tubulin. The resulting bands were quantified. Fractions 6-8, which contained the majority of the proteins, were pooled (peak #) and immunoprecipitated with protein A beads coated with polyclonal antibody against γ-tubulin or preimmune serum from the same animal (preim.). Immunoblotting of the precipitates with antibodies against NEDD1 and γ-tubulin shows that NEDD1 is specifically associated with γ-tubulin in the purified fractions.