Identification of Drosophila Dgt7, Dgt8, and Dgt9 as augmin subunits. (A) Dim γ-tubulin and MT intensities within the spindle after RNAi knockdown of Dgt7/CG2213, Dgt8/CG13879, and Dgt9/CG13914. The density of γ-tubulin or MTs inside the spindle decreased after RNAi of Dgt6–9 (n ≥ 5 each). The ratio of signal intensity in spindle (location at “b” in the lower image) to that in centrosome (“a”), which was normalized to control, is shown (±SEM). The normalized ratio for γ-tubulin and MT after Dgt7–9 knockdowns were significantly (P < 0.004) lower than in control. (B) Dgt7-HA, Flag-Dgt8, and Dgt9-GFP proteins coimmunoprecipitate with endogenous Dgt6. The supernatant (S) and precipitated (P: 40× volume of S) fractions after immunoprecipitation with anti-HA, anti-Flag, and anti-GFP antibodies were immunoblotted. (C) Dgt7-GFP, Dgt9-GFP, and Flag-Dgt8 were localized to spindle MTs during metaphase. Flag-Dgt8 was stained by anti-Flag antibody. Blue, DAPI. (Scale bars, 5 μm.)