Phenotypic analysis of the IMC formation in dd-Rab11B_N125I expressing parasites.

(A) ddRab11BN125I parasites co-expressing YFP-MyoA (green) were treated without and with 1 µM Shld-1 for 24 h. For detection of the dominant negative Rab11B, parasites were labelled with anti-myc (red). (B) ddRab11BN125I parasites co-expressing different YFP-tagged components of the glideosome/IMC (green) were treated without and with Shld-1 for 24 h. In absence of Shld-1 Gap50 can be identified in the IMC of first- and premature second-generation daughter parasites, while Gap45 can only be detected in the mature IMC (upper panel). In presence of Shld-1, early and late components appear comparably affected during IMC assembly. Initial nucleation of the daughter IMC can still take place (white arrow, second panel). MLC1 remains restricted to the pellicle of the first generation mother cell, while only a faint staining of IMC 1 can be detected (white arrow, middle panel). MLC 1 and MyoA only co-localise at the IMC of the mother, while MLC1 also accumulates close to the nucleus of forming daughter cells (white arrow, last panel). Scale bars represent 5 µm.