Figure 1. Wild type GPI-PLC and GPI-PLC-eYFP hydrolyse the VSG GPI-anchor at similar rates.

(A) Cell lines modified to contain a single copy of the wild type GPI-PLC gene (GPI-PLC/−) or a single copy modified with a C-terminal eYFP tag (GPI-PLC-eYFP/−) were lysed with 0.5% triton X-100 and samples removed over a time course. The appearance of the CRD epitope in the lysates was used as a measure of the product of GPI-PLC action; anti-BiP was used as a loading control. 2×10⁶ cell equivalents were loaded in each track. (B) The same cell lines were hypotonically lysed on ice for 5 minutes and then warmed to 37°C. Samples were taken over this time course and centrifuged to pellet cell bodies. Pellet and supernatant fractions were then analysed by Coomassie stained gels and Western blots probed with anti-CRD. 1×10⁶ cell equivalents were loaded in each track.