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. 2021 Feb 19;134(4):jcs258356. doi: 10.1242/jcs.258356

Fig. 5.

Fig. 5.

TPX2 is required for astral microtubule formation. (A) Endogenous TPX2 localizes to astral microtubules and spindle microtubules. SV589 cells in prometa/metaphase and anaphase were stained for TPX2 (green), α-tubulin (red) and DNA (blue). Panels on the right show the TPX2 signal in inverted grayscale. (B) AurkinA is a small molecule inhibitor that specifically blocks the interaction of TPX2 with Aurora A kinase (MT, microtubule). (C) AurkinA alters spindle organization. NRK cells arrested at metaphase using MG132 for 1 h were treated with AurkinA for 0, 15 or 30 min and then fixed and stained for α-tubulin. (D) AurkinA compromises astral microtubules. After 15 min of AurkinA treatment, NRK cells were fixed and stained for α-tubulin. Enlargements of the astral microtubule area are shown in the panels on the right. (E) Quantitation of the experiment shown in D. n>30, ***P<0.0001 (Student's t-test). (F) AurkinA inhibits astral microtubule growth. After 15 min of Aurkin A treatment, NRK cells were fixed and stained for EB1. Enlargements of the astral microtubule area are shown in the panels on the right. (G) Quantitation of the experiment shown in F. n>30. ***P<0.0001 (Student's t-test). In E,G, data are presented as mean±s.e.m. Maximum intensity projections of confocal z-sections are shown. Scale bars: 5 µm.