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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 4. Half-life of GPI-PLCp in Blood stream T. brucei.

Figure 4

(A), T. brucei 427 was pulsed with [35S]Met at 37°C and chased in HM1-9 medium for 6 h. Parasites were harvested at 1 h intervals during the “chase”. GPI-PLCp (from 3.5 × 106 parasites) was immunoprecipitated and analyzed by 12% SDS-PAGE followed by phosphorimaging. (The upper band in Fig. 6A was due to non-specific adsorption to the antibody since it is not detectable when stringent wash conditions are employed after immunoprecipitation of GPI-PLCp (see Fig. 4A)). (B), T. brucei 427 bloodstream form was treated with 1.2 mM cycloheximide, and then incubated at 37°C for varying amounts of time. Parasites were lysed hypotonically and assayed for GPI-PLC activity.