Fig. 1

PyDMT1 resides in the vacuole and could functionally rescue the iron defect of △smf3 yeast. a Phylogenetic analyses of Plasmodium DMT1 proteins with human and yeast (S. cerevisiae) homologs. The phylogenetic tree was generated with MEGA (Molecular Evolutionary Genetics Analysis). All Plasmodium species examined contain one DMT1 homolog. b Intraparasitic PyDMT1 localizes to Hz-containing DVs in P. yoelii. Shown here are a merge of DIC, Hoechst 33342 nuclear stain (blue), and the signal of tagged PyDMT1(green). c Indirect immunofluorescence assays of PyDMT1-HA and PyCRT-FLAG in the blood stage. The endogenous PyDMT1 gene fused to HA and the food vacuole marker PyCRT-FLAG were imaged. Shown are the HA channel (red, first row), the FLAG channel (green, second row), and a merge of both signals (third row). d Complementation of △smf3 yeast iron deficiency with PyDMT1 and an N-terminal truncated PyDMT1. The regulatory region of FET3 (− 300 bp–6 bp) directed LacZ activity (to indicate FET3 expression response). △smf3 and the parent strain by4742 transformed with the empty vector pYES2-ADH were the controls. Shown are mean values ± SD. ***P < 0.001; one-way ANOVA and Tukey’s multiple comparison test. N = 3. e Whole-cell iron content in △smf3 yeast expressing PyDMT1. Shown are mean values ± SD. *P < 0.05; t-test. N = 3. f Iron content of the vacuole from △smf3 yeast expressing PyDMT1yeast. Shown are mean values ± SD. *P < 0.05; one-way ANOVA t-test. N = 3. g Growth complementation of △smf3 yeast with the expression of PyDMT1 or an N-terminal truncated PyDMT1