Figure 5.

Ran regulates augmin in Xenopus egg extract.A (top), schematic of immunoprecipitation (IP) strategy where antibodies were conjugated to magnetic beads (top). Bottom, Western blot of IPs for a control antibody (immunoglobulin [IgG]) and antiaugmin in the presence or the absence of Ran^Q69L. B, TIRF images of augmin in Xenopus egg extract in the absence (left column) and presence (right column) of Ran^Q69L. Branching MT nucleation reactions were carried out with Alexa568-labeled tubulin and fixed after 15 min. Endogenous augmin was detected via indirect immunofluorescence with custom primary antibodies against Haus8 and Alexa488-conjugated secondary antibodies. Scale bar corresponds to 5 μm. MT, microtubule; TIRF, total internal reflection fluorescence.