Localisation of selected T. brucei clathrin-associated proteins. C-terminal triple HA tags were added to selected clathrin-associated proteins at endogenous genomic loci via homologous recombination in bloodstream-stage parasites. (A) Immunofluorescence colocalisation showed correlation between clathrin-associated proteins (red, left panels) and CHC (green, central panels). Note that TbAAKL (i), TbCAP80 (ii) and TbCAP141 (iii) showed preferential colocalisation with clathrin punctae in the vicinity of the flagellar pocket. DAPI staining is in blue. N, nucleus; K, kinetoplast (indicated on top panel only). Scale bar: 2 μm. (B) Super-resolution SIM shows close association of clathrin-associated proteins (red, left panels, anti-HA antibody) with the flagellar pocket and clathrin-positive structures (green, centre panels, anti-TbCHC antibody) close to the kinetoplast. Boxes below the main panels show z-plane resections along the axis denoted by the black arrowhead for each image. Scale bar: 500 nm.