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. 2014 Jul 8;9(7):e101471. doi: 10.1371/journal.pone.0101471

Figure 3. Identification of other augmin subunits that co-precipitate with Aug6.

Figure 3

(A) List of putative augmin subunit proteins co-precipitated with Aug6-GFP. The numbers of peptides identified in the Aug6-GFP immunoprecipitants are shown. No homology to known augmin subunits was found for AN0286. (B) Co-precipitation of Aug6-GFP, but not γ-tubulin, with Aug3-HA. Asynchronous and metaphase-accumulated cell cultures were used. Aug3-HA was immunoprecipitated by the anti-HA antibody, followed by immunoblotting of Aug6-GFP or γ-tubulin. Asterisk indicates cross-reaction of the antibody with the IgG proteins. Sup; proteins unbound to the anti-HA beads, Beads; immunoprecipitants. Equal amounts of Input and Sup were loaded, whereas the immunoprecipitants were 19- (Aug6-GFP, Aug3-HA) or 150 (γ-tubulin)-fold concentrated. (C) Amino acid sequence alignment of Aug1, Aug2, Aug3, and Aug4. Sequences from the red bread mould Neurospora crassa (Nc), Drosophila melanogaster (Dm), Homo sapiens (Hs), and the moss Physcomitrella patens (Pp) are aligned. (D) Co-localisation of Aug1–4 and AN0286 with Aug6 in metaphase. mCherry-tagged Aug1, Aug2, Aug3, Aug4, or AN0286 (purple) was co-imaged with Aug6-GFP (green) using a spinning-disc confocal microscope. See also Movie S2. Bar, 2 µm.