Figure 3.

LAMP1 is shed from the PVM towards the TVN of the parasite. (a) A lysosome is represented with LAMP1 in its membrane. The N-terminus resides in the lysosomal lumen and the C-terminus points into the cytoplasm. Therefore, mEos3.2 faces the cytoplasm. (b) LAMP1 is represented from its N- to the C-terminus. The N-terminus contains a signal peptide (SP). mEos3.2 is fused to the C-terminus of LAMP1. (c,d) HeLa cells were transfected with BFP-LC3 plus LAMP1-mEos3.2. The transfected cells were infected with wild-type P. berghei (PbWT) and allowed to develop until 24 hpi before the imaging started. (c,d) Show different staining of exactly the same cell (c) BFP-LC3 was used to identify infected cells. The inverse images show BFP-LC3 (blue) and LAMP1-mEos3.2 (green). In the merged image, the parasite was identified by the BFP-LC3 in the parasite’s PVM. The circle depicts the PVM while the TVN is surrounded by a box. (d) LAMP1-mEos3.2, which is associated with the PVM was converted in the region of interest (ROI). The parasite is indicated by a grey circle. The ROI is only represented in the enlargement (merge). Enlargements of the indicated boxed part represented next to each image show that converted lysosomes (red) are moving from the PVM towards the TVN. The image shows one representative parasite out of 10 from three independent experiments. Scale bars are 10 µm. (e) Quantification of relative fluorescent intensity in the red channel at the PVM and TVN, respectively (N = 10; mean ± SD). The graph shows that the relative red fluorescence in the PVM decreases, while it increases in the TVN.