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Fig. 1. Expression of metacaspases. (A,B) Total cell lysates were prepared from T. brucei PCF (lane 1) and BSF (lane 2). 5x106 cell equivalents were then subjected to SDS-PAGE and transferred to PVDF membrane prior to immunoblotting with immunopurified rabbit anti-MCA2-MCA3 antibodies (A) or with sheep anti-MCA5 antiserum (B). MCA2, MCA3 and MCA5 are indicated. A crossreacting 48 kDa protein (A, see *) and an antibody against EF-1
demonstrates equal loading. (C) Western blots prepared with BSF wild-type (lane 1) and transgenic parasites expressing an ectopic copy of either MCA3HA (lane 2) or MCA2HA (lane 3) were probed with a monoclonal anti-HA antibody. MCA2HA and MCA3HA are indicated at their predicted sizes of 40 and 41 kDa, respectively.