Fig. 7: Mutating the ortholog of DegP2 in P. falciparum reduces susceptibility to DHA.

a Diagram showing strategy to knockout PfDegP in the Cam3.II^WT background. b PCR confirmation of successful KO of PfDeg2. PCR 1 (using primers P7 and 8) confirmed the loss of exons 3 and 4 in the two DegP KO clones; clone G10 was used for the remainder of this study. PCR 2 (using primers P7 and 5) demonstrated a shorter product upon successful deletion of two exons. c Ring-stage survival assay, performed as above, using the Cam3.II^WT, Cam3.II^ΔPfDegP, and the K13 mutant Cam3.II^R539T lines. Results are mean ± SEM for n = 3 independent replicates; p-values derive from two-tailed, unpaired t-tests. d Schizont-stage survival assay following a 4 h pulse of 700 nM DHA. Results are mean ± SEM for n = 4 independent replicates; p-values derived from two-tailed unpaired t-test. e–g Heme measured from the free (e), hemoglobin-associated (f), and hemozoin (g) pools from trophozoites for various strains. Heme concentrations were calibrated to a standard curve and normalized to the number of parasites per sample to calculate fg/cell. Results are mean ± SD for n = 4 independent replicates; p-values from one-way ANOVA.