Fig 2. TbUsp7 but not TbVdu1 knockdown perturbs endocytosis.

(A) Knockdown of TbUsp7 and TbVdu1 in the presence (+, open bar) or absence (-, closed bar) of RNAi induction as measured by qRT-PCR. Results were normalised to β-tubulin and error bars denote the standard error of the mean (n = 3). (B) Phase contrast images showing the presence or absence of the BigEye phenotype under TbUsp7 and TbVdu1 knockdown. The enlarged flagellar pocket is indicated by a black arrowhead. Scale bar = 2.5 μm. (C) One hundred cells were analysed from uninduced or induced cultures of TbVdu1 (open bar) or TbUsp7 (closed bar) and scored for the appearance of an enlarged flagellar pocket, manifest as a phase-light vacuole at the posterior end of the cell. Results represent an average of four independent experiments with error bars denoting the standard error. (D) ConA uptake in the presence and absence of TbUsp7 knockdown. Following induction of RNAi, cells were incubated with FITC-conjugated ConA (green). Note that only an example of a cell with an enlarged flagellar pocket is presented for TbUsp7. Induced cells that do not display this morphology have normal ConA uptake. Scale bar = 2.0 μm. All cells were co-stained with DAPI (blue) to visualise nuclear and kinetoplast DNA.