Journal: Nature Communications

Article Title: A G358S mutation in the Plasmodium falciparum Na + pump PfATP4 confers clinically-relevant resistance to cipargamin

doi: 10.1038/s41467-022-33403-9

Figure Lengend Snippet: IC 50 values are shown for cipargamin ( a–c ), (+)-SJ733 ( d – f ) and dihydroartemisinin ( g – i ) against HCR parasites (clone #1 in red and clone #2 in dark red) generated through in vitro evolution with the Dd2-PfATP4 T418N,P990R line (grey; with data for its Dd2 parent also shown in black; a , d , g , circles), Dd2-Polδ-PfATP4 G358S (red) parasites generated through in vitro evolution with the Dd2-Polδ line (black; b , e , h , triangles), and two NF54 G358S lines (1 in red and 2 in dark red) generated using CRISPR-Cas9 with the NF54 WT line (black; along with the matched recombinant control line NF54 CTL in black; ( c , f , i ), upside down triangles). Within each panel, all lines were tested in parallel in each experiment. The symbols show the data from individual experiments and the bars show the mean + SEM. The data are from the following number of independent experiments, performed on different days: 16 ( a ), 7 ( b ), 4 ( c ), 5 ( d ), 4 ( e ), 4 ( f ), 4 ( g ), 4 ( h ) and 4 ( i ). For each compound, the IC 50 values for each cipargamin-resistant line were compared with those of its direct parent using two-tailed paired t tests ( P values ≤ 0.05 indicate statistical significance and are shown in bold). Source data are provided as a Source Data file.

Article Snippet: The transmission-competent NF54 WT strain (BEI Resources, MRA-1000) provided by Johns Hopkins University was used for pfatp4 gene editing.

Techniques: Generated, In Vitro, CRISPR, Recombinant, Control, Two Tailed Test

Journal: Nature Communications

Article Title: A G358S mutation in the Plasmodium falciparum Na + pump PfATP4 confers clinically-relevant resistance to cipargamin

doi: 10.1038/s41467-022-33403-9

Figure Lengend Snippet: a Left Panel: Oocyst numbers in the midguts of Anopheles stephensi mosquitoes fed on gametocyte cultures of the parasite lines indicated ( n = 23–26 mosquitoes per line; determined on Day 12 post blood meal). The red bar indicates the mean. Data for two NF54 WT cultures (prepared from separate vials that had been frozen on different dates; closed circles) are shown. Right Panel: Pooled data for statistical analysis. Because of the high variability inherent in these data, we pooled the two NF54 WT controls (closed circles) and the three mutant lines (NF54 G358S-1 , -2 , and -3 ; closed triangles) for analysis. There was a small difference in oocyst numbers between NF54 WT and NF54 G358S ( P = 0.04; Mann–Whitney test, two-tailed). There was a larger difference in oocyst numbers between NF54 WT and NF54 CTL (open squares; P = 0.0007; Mann–Whitney test, two-tailed). P values ≤ 0.05 indicate statistical significance and are shown in bold. b Numbers of salivary gland sporozoites, determined on Day 16 post blood meal. The data are the mean number of sporozoites per mosquito. For each parasite line, 20 mosquitoes were dissected, their salivary glands pooled and homogenised, and sporozoites counted using a haemocytometer. Source data are provided as a Source Data file.

Article Snippet: The transmission-competent NF54 WT strain (BEI Resources, MRA-1000) provided by Johns Hopkins University was used for pfatp4 gene editing.

Techniques: Mutagenesis, MANN-WHITNEY, Two Tailed Test