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. Author manuscript; available in PMC: 2011 Jul 1.
Published in final edited form as: Exp Parasitol. 2010 Jan 28;125(3):222–229. doi: 10.1016/j.exppara.2010.01.014

Figure 2. Early-stage Procyclic T. brucei 427 Retain GPI-PLCp.

Figure 2

Culture-adapted T. brucei 427 (CA-427) was resuspended in SDM-79 transformation medium (3.3 × 106/ml) and incubated at 27°C. Parasites (2 × 107) were harvested for analysis as described in the legend to Fig. 1. (A), Total GPI-PLCp activity (per 107 cells), expressed as a proportion of the activity of day zero cells. (B), Changes in protein expression. Cells (2 × 107) were withdrawn at the indicated time points, lysed hypotonically, and membrane proteins resolved by SDS-PAGE (12%, minigel). VSG, procyclin (PARP) and tubulin were analyzed as described in legend to Fig. 1C PCF, established procyclic; Day #, days post initiation of differentiation; TUB, tubulin, PARP, procyclin. (C), Analysis of procyclin and VSG Expression by Flow Cytometry. Cell surface expression of VSG and procyclin in differentiating T. brucei 427 was determined by flow cytometry (detailed in Materials and Methods).