PfSPM3 is dispensable for the asexual replication cycle. (A) Representative live-cell microscopy of 3D7-PfSPM3-GFP-glmS schizonts cultured either with or without (control) 2.5 mM GLCN at 40 h after addition of GLCN. Scale bar, 2 μm. (B) Growth of 3D7 and 3D7-PfSPM3-GFP-glmS parasites treated with or without 2.5 mM GLCN after two parasite replication cycles, as determined by flow cytometry. Shown are relative parasitemia values, which were obtained by dividing the parasitemia of GLCN-treated cultures by the parasitemia of the corresponding untreated ones. Values are means and standard deviations (SD) from independent growth experiments, with the number of experiments (n) indicated. P values were determined with a two-tailed unpaired t test with Welch’s correction. (C) Localization of truncated PfSPM3-TGD-GFP fusion protein by live-cell microscopy across the intraerythrocytic development in the 3D7-iGP background. (D) Growth curves of 3D7-iGP-PfSPM3-TGD versus 3D7-iGP parasites after two parasite replication cycles, as determined by flow cytometry. Three independent growth experiments were performed, and a summary is shown as percentage of growth compared to the parental parasite line. Values are means and SD from independent growth experiments, with the number of experiments (n) indicated. P values were determined with a one-sample t test. (E) Colocalization of the truncated PfSPM3-TGD-GFP fusion protein with TubulinTracker and the IMC marker proteins ALV5mCherry and PF3D7_ 1345600mCherry. Nuclei were stained with Hoechst 33342. Bar, 2 μm. Zoom factor, 400%, with a scale bar of 1 μm. Wichers-Misterek JS, Binder AM, Mesén-Ramírez P, Dorner LP, Safavi S, Fuchs G, Lenz TL, Bachmann A, Wilson D, Frischknecht F, Gilberger TW. A Microtubule-Associated Protein Is Essential for Malaria Parasite Transmission. mBio. 2023.