Figure 1. Wac is an integral component... | Rockefeller University Press

Figure 1.

$Figure 1. Wac is an integral component of Augmin. (A) Wac and Dgt2 reciprocally coimmunoprecipitated. Wac or Dgt2 was immunoprecipitated from the soluble fraction (input) of S2 cell extract using preimmune sera (PreI) as a control (Ctrl) and immunoblotted for these two proteins. > and − indicate 16.5 kD and 25 kD, respectively. (B) Decreased amounts of Wac and Dgt2 after depletion of Augmin subunits (Dgt2–6). (C) The frequencies of monopolar spindles and abnormal bipolar spindles, typically with reduced microtubule density, after 5 d of dsRNA treatment. (D) Time sequences of mitotic progression in S2 cells expressing GFP–α-tubulin after 3 d of dsRNA incubation. Time (minutes:seconds) after the nuclear envelope breakdown is shown. (E) Change in the GFP–α-tubulin signal intensity on the spindles shown in B and change in the spindle length. The intensity was normalized against signals around the poles at prophase. The arrowheads indicate anaphase onset. (F) Typical spindle morphologies of single centrosomin (Cnn) depletion and of codepletion with Wac. (G) The γ- and α-tubulin signal intensity on the spindle relative to the poles after RNAi. IP, immunoprecipitation. Error bars indicate SD. Bars, 10 µm.$

Wac is an integral component of Augmin. (A) Wac and Dgt2 reciprocally coimmunoprecipitated. Wac or Dgt2 was immunoprecipitated from the soluble fraction (input) of S2 cell extract using preimmune sera (PreI) as a control (Ctrl) and immunoblotted for these two proteins. > and − indicate 16.5 kD and 25 kD, respectively. (B) Decreased amounts of Wac and Dgt2 after depletion of Augmin subunits (Dgt2–6). (C) The frequencies of monopolar spindles and abnormal bipolar spindles, typically with reduced microtubule density, after 5 d of dsRNA treatment. (D) Time sequences of mitotic progression in S2 cells expressing GFP–α-tubulin after 3 d of dsRNA incubation. Time (minutes:seconds) after the nuclear envelope breakdown is shown. (E) Change in the GFP–α-tubulin signal intensity on the spindles shown in B and change in the spindle length. The intensity was normalized against signals around the poles at prophase. The arrowheads indicate anaphase onset. (F) Typical spindle morphologies of single centrosomin (Cnn) depletion and of codepletion with Wac. (G) The γ- and α-tubulin signal intensity on the spindle relative to the poles after RNAi. IP, immunoprecipitation. Error bars indicate SD. Bars, 10 µm.