Fig. 3: PfAnchor is required for the release of segregated merozoites but dispensable for egress.

a Growth analysis of PfAnchor-expressing and deficient parasites over two replication cycles. Parasitemia was quantified by light microscopy with three biological replicates. Hemacolor-stained smears show abnormal late-stage morphology prior to egress and no ring-stage parasites after egress in PfAnchor-deficient cultures. Representative growth curve from one of the three independent experiments with technical replicates. Data are presented as mean ± SD. Scale bar 5 µm. b Representative live microscopy images of PfAnchor-expressing (top) and PfAnchor-deficient (bottom) parasites at egress. While PfAnchor-expressing parasites release individual merozoites, PfAnchor-deficient parasites form clumps of daughter cells that remain tethered (abnormal morphology). Scale bar 5 µm. c Quantification of normal and abnormal daughter parasite morphologies after egress from live microscopy experiments. PfAnchor-expressing parasites had 89.3 ± 9.5% normal morphology, whereas PfAnchor-deficient parasites showed 10.7 ± 9.3% normal morphology. Unpaired t-tests with error bars indicating mean ± SD from (+ aTC: 61 cells over 3 replicates, -aTC 39 cells over 3 replicates). d Representative live microscopy images from one of the three independent experiments of PfAnchor-expressing (top) and PfAnchor-deficient (bottom) parasites at egress, with the apicoplast labeled in green. Parasites expressing PfAnchor show daughters moving apart with the apicoplast labeled in green (ACP-GFP). In PfAnchor-expressing parasites, daughter cells separate and properly inherit apicoplast fragments. In contrast, in PfAnchor-deficient parasites, daughters remain clumped with persistent apicoplast material across the cluster or retained in the residual body. Scale bar 5 µm. Source data are provided as a Source Data file.