Fig. 4. Location of metacaspases. (A) BSF T. brucei parasites expressing either MCA2HA or MCA3HA proteins were fixed and co-stained with anti-HA monoclonal antibody (red) and sheep anti-MCA5 antiserum (green). (B) BSF parasites were fixed and stained with rabbit anti-RAB11 (red) and sheep anti-MCA5 antiserum (green). Single-slice images were analysed with volume deconvolution by using nearest-neighbour algorithm. (C) BSFs were incubated with rabbit anti-VSG221 antiserum at 4°C for 30 minutes and then returned to 37°C for 30 minutes. Fixed and permeabilised cells were stained with anti-MCA5 antiserum (green) and anti-rabbit IgG (red). For all images DNA was visualised with DAPI (blue). DIC images of the parasite are shown. Colocalisation is shown in yellow. Bars, 10 µm.