1. Quantification of drug cross-resistance between mutants selected for I-BET151 resistance and F0002 resistance.
Quantification of drug cross-resistance between mutants selected for I-BET151 resistance and F0002 resistance. (A) Chemical structures of F (F0002) and I-BET151. PubChem identifiers and other properties relevant to their drug-like properties are listed. (B) Three F0002-resistant mutants were challenged in in vitro growth assays with 3.13 to 100 μM I-BET151, and the effect on parasite growth was assessed by direct parasite counting. All 3 mutants shown had significantly different dose-response curves (Table 2) and increases in IC50 values compared to wild-type assays performed in parallel (P < 0.05) (Table 2). All wild-type (filled shapes, dashed gray lines) and mutant (open shapes, black lines) fitted curves are shown and are paired by shape. n = 1 for each drug concentration for each mutant. IC50 and P values are listed in Table 2. (C) Six I-BET151-resistant mutants were challenged in identical assays with to 10 μM F0002, and the effect on parasite growth was assessed by direct parasite counting. The 3 mutants shown (out of 6 total) had significantly different dose-response curves (P < 0.05) (Table 2) compared to wild type. Shape, line, and color are as in panel B. n = 1 for each drug concentration for each mutant. IC50 and P values are listed in Table 2. (D and E) Two TgPRELID F0002-resistant mutant clones generated by CRISPR/CAS9 and nonhomologous end-joining DNA repair were further tested for their resistance to 50 μM I-BET151 (E) and 10 μM F0002 (D) by direct parasite counting 24 h after drug exposure. Following 2-way ANOVA, all values were compared to RH with vehicle using post hoc tests as described in Materials and Methods. *, P < 0.05; **, P < 0.01, compared to RH with vehicle. n = 2 wells per strain-drug combination.
Victoria Jeffers et al. mSphere 2017; doi: /mSphere