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BioMed Research International
Volume 2014, Article ID 893272, 12 pages
http://dx.doi.org/10.1155/2014/893272
Research Article

Tracking the Biogenesis and Inheritance of Subpellicular Microtubule in Trypanosoma brucei with Inducible YFP-α-Tubulin

1Department of Biological Sciences, National University of Singapore, Singapore 117543
2Centre for BioImaging Sciences, National University of Singapore, Singapore 117546

Received 27 December 2013; Accepted 19 February 2014; Published 30 March 2014

Academic Editor: Wanderley de Souza

Copyright © 2014 Omar Sheriff et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

Figure S1: YFP-EB1 localization during T. brucei cell cycle. Cells stably expressing YFP-EB1 (green) were fixed with cold methanol and labeled for FAZ (red) and DNA (blue). Whereas YFP-EB1 was most readily detected at the posterior tip of the cells, YFP-EB1 was occasionally found along the new FAZ. YFP-EB1 labelling at the posterior tip varied with cell cycle progression, forming a dot most of the time (A, B, C), elongating into a line at mitosis (D), and reforming two separate dots at the posterior tips of the two daughters at cell division (E). Arrows, YFP-EB1 staining at the posterior tip of the cell; double headed arrow: elongated YFP-EB1 pattern during mitosis; white lines: occassional YFP-EB1 labelling near the new FAZ.

Figure S2: Characterization of anti-EB1. Cell lysates from 29.13 control and YFP-EB1 stable cells were fractionated on SDS-PAGE and immunoprobed with anti-EB1 or anti-GFP. Anti-EB1 reacted to a single ~57 KDa band corresponding to the estimated size of T. brucei EB1 in wild type 29.13 cell lysates. An additional ~84 KDa band was detected in YFP-EB1 cell lysates and this band was also detected by anti-GFP.

  1. Supplementary Figure S1
  2. Supplementary Figure S2