Characterization of Tb 11.NT.29 . (A) Sequence conservation of Tb11.NT.29 across kinetoplastids. Amino acid sequences from T. brucei (Tbr), T. cruzi (Tcr) and L. major (Lm) were aligned with ClustalW and conserved residues are shaded with black boxes while similar residues are shaded in grey. The predicted transmembrane domain (TM helix) is indicated. (B) Immunofluorescence analysis on cells expressing a C-terminal HA-tagged version of Tb11.NT.29. Permeabilized cells (left three panels) and non-permeabilized cells (right three panels) were probed for HA (green) and BiP (red) and DNA was stained with Hoechst (blue). (C) Immunofluorescence analysis of procyclin as described in panel (B). (D) Cell fractionation of parasites expressing a GFP-tagged version of Tb11.NT.29. Western blot analysis was performed against GFP-tagged Tb11.NT.29 (top panel), procyclin (second from top), BiP (third panel from top) and HSP 70 (bottom panel) on total (T), cytoplasmic (C), membrane (M), nuclear (N) and cytoskeleton (CSK) fractions.