Knockdown of PfPDK1 expression in NF54/PDK1 cKD parasites has no major effect on asexual parasite growth. (A) Expression of PfPDK1-GFPDD in ring (18 to 24 hpi), trophozoite (24 to 30 hpi), and schizont (42 to 48 hpi) stages cultured under protein-stabilising (+Shield-1) conditions by live cell fluorescence imaging. Representative fluorescence images are shown. Parasite DNA was stained with Hoechst. Scale bar = 5 μm. PfPDK1-GFPDD is expressed in the cytosol and nucleus throughout the IDC with highest expression in schizonts(B) Expression of PfPDK1-GFPDD under protein-depleting (–Shield-1) and control conditions (+Shield-1) by live cell fluorescence imaging and western blot analysis. Synchronous parasites (0 to 8 hpi) were split (±Shield-1) 40 hours before collection of the samples. PfPDK1-GFPDD expression was efficiently reduced after Shield-1 removal compared to parasites grown in the presence of Shield-1Representative fluorescence images are shown. Parasite DNA was stained with Hoechst. Scale bar = 5 μm.
Hitz E, Wiedemar N, Passecker A, Graça BAS, Scheurer C, Wittlin S, Brancucci NMB, Vakonakis I, Mäser P, Voss TS. The 3-phosphoinositide-dependent protein kinase 1 is an essential upstream activator of protein kinase A in malaria parasites. PLoS Biol. 2021 Dec 8;19(12):e3001483.