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. 2016 May 23;213(4):425–433. doi: 10.1083/jcb.201603050

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© 2016 Pamula et al.

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Figure 3. — Design and characterization of chimeric β tubulin constructs. (A) Design of tail-swapped β tubulin constructs, with amino acid sequence derived from α/βIIB (black) and α/βIII (gray). (B) Schematic of tubulin heterodimer indicating β tubulin C-terminal tail (black). Amino acid sequences from the C terminus of βIIB and βIII are shown. (C) SDS-PAGE analysis (1, nickel affinity elution; 2, TOG affinity elution; Coomassie stain). (D) Western blot (WB) analysis. Full blots are provided in Fig. S2 A. (E) Protein elution profiles from size-exclusion chromatography. Peak volume: 14.2 ml (α/βIIB-tail-III); 14.1 ml (α/βIII-tail-IIB). Void volume (V₀) is 7 ml. A.U., arbitrary units. (F and G) TIRF images of taxol-stabilized (F) or GMPCPP (G) microtubules. Bars, 3 µm.