Fig. 2: PfMYST knockdown or inhibition leads to ring-stage survival and recrudescence in P. falciparum.

a Correlation analysis between the transcription levels of Pfmyst and ART susceptibility in field-adapted parasites (n = 28) carrying the WT PfK13 gene. Parasites were exposed to 50 nM DHA and then cultured normally for 66 hours. The correlation analysis was performed using GraphPad Prism 8.0 software, and data were presented as mean value from three independent experiments with technical triplicates. b Schematic illustrating the CRISPR/Cas9-mediated fusion of the glmS sequence and Ty1 tag at the 3’ terminus of PfMYST, resulting in the PfM-KD parasite line (Pfmyst-ty1-glmS). The fusion event was verified by PCR with gDNA extracted from the transfectant. Similar trends were observed in two independent experiments. c Intraerythrocytic growth curves of two subclones (C4, D5) of the Pfmyst-ty1-glmS line and the WT 3D7 control, with and without 5 mM GlcN treatment for 6 h. Data were presented as mean ± SEM from three independent replicates with technical triplicates. d Western Blot analysis of the Pfmyst-ty1-glmS line and WT 3D7 parasites, in which parasites were pretreated with 5 mM GlcN for 6 h and total protein extracts were obtained from ring, trophozoite, and schizont-stage parasites. The analysis utilized a commercial antibody against the Ty1 epitope, alongside Aldolase as the internal control. Similar trends were observed in two independent experiments; RT-qPCR analysis of Pfmyst transcript level at ring (0-6 hpi), trophozoite (20–26 hpi), and schizont-stage (36–42 hpi) with/without the exposure of GlcN. Data were presented as mean ± SEM from four independent experiments with technical triplicates. P-values were determined using the two-tailed student’s t test. e RSA with DHA concentrations ranging from 50 nM to 700 nM was tested on different parasites, including PfM-KD parasites with or without GlcN treatment, WT and Rrp6 knockdown parasites (Pfrrp6-Ty1-glmS). Data were presented as mean ± SEM from four independent experiments with technical duplicates. f The effects of histone acetyltransferase inhibitors on parasite RSA_0-3h induced by DHA. WT parasites were pre-incubated with PfMYST specific inhibitor (NU9056) and PfGCN5 inhibitors (curcumin, and anacardic acid) for 12 h before RSA_0-3h. Various DHA concentrations (50 nM, 200 nM, 700 nM) were tested. Data were presented as mean ± SEM from three independent experiments with technical triplicates. P-values were determined using One-way ANOVA with Bonferroni correction compared with vehicle control. CCM: curcumin, AA: anacardic acid. Source data are provided as a Source Data file.