GlcN-induced knockdown of PfHsp70x does not affect intraerythrocytic growth. GlcN-induced knockdown of PfHsp70x does not affect intraerythrocytic growth. (A) PfHsp70x-glmS, PfHsp70x-M9, and 3D7 (parental) cell lines were seeded at equal parasitemia in triplicate and grown in normal culturing medium. Parasitemia was measured every 24 h using flow cytometry. Data are fit to an exponential growth equation and are represented as means ± standard errors of the means (SEM) (error bars) (n = 3). (B and C) PfHsp70x-glmS and PfHsp70x-M9 parasites were seeded at equal parasitemia in triplicate. Cultures were grown in the presence of either 5 mM or 10 mM GlcN. Parasitemia was measured every 24 h using flow cytometry. Data are fit to an exponential growth equation and are represented as means ± SEM (n = 3). (D) PfHsp70x-glmS and PfHsp70x-M9 parasites were grown in the presence of 7.5 mM GlcN. Schizont-stage parasites were purified on a Percoll gradient every 24 h, and whole-parasite lysates were used for Western blot analysis. The membrane was probed with anti-HA (α-HA) and anti-PfEF1α (loading control) antibodies. The positions of molecular mass markers (in kilodaltons) are indicated to the left of the blot. David W. Cobb et al. mSphere 2017; doi:10.1128/mSphere.00363-17