FIG 3.

mRNA levels for selected genes associated with HePC resistance and effect of energy depletion on HePC uptake. (A) qRT-PCR experiments carried out on LdR30.1 parasites showed decreased HePC transporter expression (LdMT/LdRos3 subunits), as well as reduced transcription of the ABC transporters MDR1 and ABCG4, compared to the WT strain, using the GAPDH gene as a housekeeping gene. Similar results were obtained when using the SOD gene. Samples were analyzed in triplicate, and the error bars indicate standard deviations (*, P < 0.05). (B) LdR30.1 and WT parasites were incubated with 5 μM HePC-BODIPY with (solid histograms) or without (open histograms) KCN to deplete intracellular ATP pools. Drug intake was followed by flow cytometry. The excitation wavelength was set at 488 nm, and emission was detected in the FL1 channel. Parasite autofluorescence (broken profiles on the left) is also shown. A total of 10,000 cells were counted for each histogram. Experiments were done in triplicate with profiles similar to those shown.