PfDyn2 localizes to the apicoplast and mitochondria, and is essential for growth and replication. (A) Super-resolution microscopy showing 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 α-PyACP (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 α-PfGAP45. PfDyn2, mitochondria, and nuclei were labeled as shown in (A). (C) U-ExM of PfDyn2-3HAapt parasites showing localization of PfDyn2 in mid- and late schizogony. PfDyn2, the apicoplast, and mitochondria were labeled as shown in (A). Proteins were labeled by AlexaFluor 405 NHS-Ester and shown in gray. White boxes indicate regions zoomed-in below the larger images; white arrows highlight PfDyn2 encircling the branches of the apicoplast or mitochondrion. (D) Immunoblot showing extent of PfDyn2 knockdown over 36 hours 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 hours (two replicative cycles). Data shown as mean ± SD (n = 3). All scale bars = 1 µm, except for the non-inset images in (C) where scale bars = 5 µm. Morano AA, Xu W, Navarro FM, Shadija N, Dvorin JD, Ke H. The dynamin-related protein PfDyn2 is essential for both apicoplast and mitochondrial fission in Plasmodium falciparum. mBio. 2024 29:e0303624. PMID: 39611847.