Figure 5 | Scientific Reports

Figure 5

From: A Plasmodium plasma membrane reporter reveals membrane dynamics by live-cell microscopy

Figure 5

FLIP experiments show that early membrane accumulations are connected to the surrounding PPM. HeLa cells were infected with PbPMP1-GFP parasites and a possible connection of membrane accumulations to the surrounding PPM was analyzed by FLIP experiments at 48 hpi. In each FLIP experiment, a single region of the surrounding PPM (white square) was repeatedly bleached and the fluorescence intensity of a membrane accumulation (Acc, red square) and another region on the surrounding plasma membrane (Mem, green square) was determined. As a control (Con), bleaching was performed outside of the parasite (white square) and the fluorescence intensity of the whole parasite was measured (blue circle). For all experiments, an image was acquired before bleaching and the corresponding fluorescence intensity was set to 100%. (a) Representative stills from a FLIP and a control parasite, with bleaching cycles indicated in the upper right. Pre = prebleaching. (b) Fluorescence intensity over time. (c) Statistical evaluation of the loss of fluorescence after 40 bleaching cycles. Shown are means with SD of 21 FLIP and 21 control parasites, which were obtained in two independent experiments. For statistical analysis, a one-way ANOVA followed by a Holm-Sidak multiple comparison test was performed (**** p < 0.0001, n.s. = not significant). (d) Schematic drawing of first membrane invaginations that presumably appear as membrane accumulations in a two-dimensional representation.