Figure 3.
NMTi IMP-1002 Shows On-Target Specificity
(A) Growth assay with P. falciparum 3D7[G386E] clones and [WT] showing shift in EC50 for both clones from transfections with independent guides (n = 3). Unpaired Welch t test not assuming equal standard deviations, p = 0.01.
(B) NMT substrates labeled with YnMyr in WT and G386E parasites after treatment with NaOH to hydrolyze the base-labile ester-linkage incorporation of YnMyr into GPI-anchored proteins. Although WT parasites show a decreased band intensity in response to the NMTi treatment, G386E parasite proteins remain unchanged compared with those of the DMSO control. The Coomassie blue-stained gel shows proportional loading of total protein for each sample.
(C) Percentage of maximum count of Hoechst-stained WT and G386E parasites used to determine the number of nuclei per sample.
(D) P. falciparum [G386E] parasites show 0% inhibition at 140 nM. Graph shows flow cytometry data of Hoechst-stained parasites in (C) treated with 140 nM IMP-1002 from 1 to 45 h post-invasion. The median fluorescent intensity (MFI) of the Hoechst-positive parasites was normalized to the MFI of parasites containing one nucleus (ring sample) in (C) to determine the average number of nuclei per sample. While WT parasites show a drop-in the number of nuclei and therefore an arrest in development at 140 nM of NMTi, G386E parasites show no arrest in development. All errors bars in this figure indicate standard deviation of the mean.