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. Author manuscript; available in PMC: 2014 Oct 14.
Published in final edited form as: Dev Cell. 2013 Sep 26;27(1):47–59. doi: 10.1016/j.devcel.2013.08.002

Figure 3. Dppa2 inhibits microtubule assembly around chromatin and in vitro.

Figure 3

(A) Dppa2 inhibits sperm aster microtubule assembly. Demembranated sperm nuclei were added together with calcium to metaphase extracts supplemented with rhodamine-labeled tubulin (red). Samples were fixed and stained with Hoechst 33342 (blue).

(B) Quantification of tubulin fluorescence intensity from (A). Data shown indicate mean and standard error from > 30 asters per sample and are representative of 3 independent experiments.

(C) Dppa2 inhibits spindle assembly in a dose-dependent manner. Metaphase spindles were assembled in extracts supplemented with MBP-Dppa2 fusion proteins and rhodamine-labeled tubulin.

(D) Quantification of spindle length from (C). Data shown are mean and standard deviation from 30 spindles per sample.

(E) Chromatin-independent microtubule assembly in Xenopus egg extracts. Recombinant GST-Dppa2 protein was added to metaphase extracts together with 0.5 % DMSO. Polymerized microtubules were recovered by pelleting and analyzed by Coomassie staining.

(F) Dppa2 inhibits microtubule polymerization in vitro. MBP-Dppa2 was added to purified bovine tubulin together with 0.5 % DMSO. Polymerized microtubules were recovered by pelleting and analyzed by Coomassie staining.

Scale bars, 10 μm.

See also Figure S3.