English: The parasite clone stably expressing ddFYVE was transfected with FNR-RFP and grown intracellularly without Shield-1 for 18 hours. Data acquisition was started 2 h after addition of 1 µM Shield-1 and images were taken every 10 minutes. The video shows an overlay of phase contrast, ddFYVE (green) and FNR-RFP (red) images. The vacuole of interest containing four parasites co-expressing both ddFYVE and FNR-RFP is located in the upper right corner at the beginning of the video. During apicoplast elongation, the FNR signal surrounds the GFP label and there is no GFP staining in the residual body (t = 120–200 min). Later, the FNR signal elongates further and moves away from GFP (t = 240 min). Seven small distinct FNR spots (one slightly out of focus) in the sub-apical part correspond to the apicoplasts of the newly formed daughter cells, the eighth one having lost the label. Additional FNR spots are found close to or co-localized with a large GFP signal that ultimately accumulates in the residual body (t = 300 min). The newly apicoplasts in the newly formed daughter cells rapidly became GFP positive (not visible in the merged images). Selected images of the vacuole of interest are depicted in Figure 2B.