PfDyn2 localizes to the apicoplast and mitochondria and is essential for growth and replication a) Super-resolution microscopy detects PfDyn2’s localization to the apicoplast and mitochondria at different stages of schizogony in PfDyn2-3HAapt parasites. PfDyn2 was probed with anti-HA. The apicoplast was probed with anti-ACP (acyl carrier protein). Mitochondria were labeled with MitoTracker Orange CMTMRos. Nuclei were stained by Hoechst. b) Super-resolution microscopy verifies PfDyn2’s localization to mitochondria at different stages of IMC development in the PfDyn2-3HAapt parasites. IMC was probed by anti-PfGAP45. PfDyn2, mitochondria, and nuclei were labelled as shown in (a). c) U-ExM of PfDyn2-3HAapt parasites comparing localization of PfDyn2 at different stages of segmentation (early, mid, and late). PfDyn2, the apicoplast, and mitochondria were labeled as shown in (a). Proteins were labeled by AlexaFluor 405 NHS-Ester and shown in grey. d) Immunoblot showing extent of PfDyn2 knockdown over 36-h following ATc removal. PfExp2, exported protein 2, was used as a loading control. e) Growth curve comparing replicative fitness of PfDyn2-sufficient (+ATc) and -deficient (-ATc) parasites over 96-h (2 replicative cycles). Parasitemia was determined by microscopy examination of Giemsa-stained thin blood smears and was the product of actual parasitemia and splitting factors. Data shown as mean ± SD (n=3). Scale bars for a and b = 1 μm, for c = 5 μm. Morano AA, Xu W, Shadija N, Dvorin JD, Ke H. The dynamin-related protein Dyn2 is essential for both apicoplast and mitochondrial fission in Plasmodium falciparum. bioRxiv [Preprint]. 2024 2024.03.15.585229. PMID: 38559241