Figure 2

Fluo-4 fluorescence in P. falciparum clones with varying PfMDR1 and PfCRT mutations. Parasites were incubated with 5 µM Fluo-4 AM in Ringer’s solution for 50 min at 37°C, then washed with Ringer’s solution and transferred onto a microscope chamber. a Single images of P. falciparum clones. Scale bar 5 µm. b Mean Fluo-4 fluorescence ratio measured from the digestive vacuole (DV) and the cytosol. When treated with tariquidar (TQ), parasites were pre-incubated with 100 nM TQ for 10 min at 37°C before adding Fluo-4 AM for 50 min. Total n ≥ 80 for each strain, done in three independent experiments on different days. Error bars represent SEM. ***p < 0.0001. c Western blot of synchronized trophozoite stage parasites using anti-PfMDR1 antibodies. Anti-PfHSP70 was used as a loading control. PfMDR1 protein expression levels were normalized to PfHSP70 and measured in triplicate ± SEM.