Journal: Journal of medicinal chemistry

Article Title: From DNA-Encoded Library Screening to AM-9747 : An MTA-Cooperative PRMT5 Inhibitor with Potent Oral In Vivo Efficacy.

doi: 10.1021/acs.jmedchem.4c03101

Figure Lengend Snippet: Figure 7. (A) AM-9747 viability in HCT116 cellular assay (blue circles, MTAP-WT; red squares MTAP-del). Viability was measured by a CellTiter-Glo assay, and cellular MTA-selectivity was determined as (HCT116-WT IC50/MTAP-del IC50). (B) HCT116-WT and MTAP-del global SDMA levels were assessed by an in-cell imaging assay after 3 days of treatment with AM-9747.

Article Snippet: Cells were then stained with an anti-SDMA polyclonal antibody (CST#13222S at 1:2000 dilution) in a wash buffer and incubated at RT for 2 h. Cells were washed with 200 mL of Wash Buffer and stained with secondary antibodies (goat antirabbit-alexa-488 Cat #A11034, Invitrogen at 1:2000 dilution) and Hoechst 33342 DNA dye (1:5000 dilution) for 1 h at RT in the dark.

Techniques: Glo Assay, Imaging

Journal: Communications biology

Article Title: A type II protein arginine methyltransferase regulates merozoite invasion in Plasmodium falciparum.

doi: 10.1038/s42003-023-05038-z

Figure Lengend Snippet: Fig. 2 Alteration of parasite growth and protein modifications upon PfPRMT5 disruption. a Growth curves of two PfPRMT5 disruptants (ΔPfPRMT5-1 and ΔPfPRMT5-2) compared to WT 3D7 parasite. The disruptants grew significantly slower than 3D7 (ANOVA, P < 0.01, n = 3 biological replicates). b The IDC progression profiles of the PfPRMT5 disruptant (ΔPfPRMT5-1) along with 3D7 control showing the proportions of the ring, trophozoite, and schizont stages through a 52 h period. The dynamics of the ring, trophozoite, and schizont stages were very similar between the 3D7 and ΔPfPRMT5-1. c Distribution of merozoite numbers in schizonts from 3D7 and two PfPRMT5 disruptants. The PfPRMT5 disruptants had fewer numbers of merozoites in schizonts than in 3D7 (ANOVA, P < 0.05, n = 3 biologically independent experiments). d The capacity of invasion by merozoites from PfPRMT5 disruptant (ΔPfPRMT5-1). The invasion rate is significantly lower than the 3D7 control (P < 0.01, n = 3 biologically independent experiments). e Western blots show a substantial decrease of H3R2me2s and relatively stable H3R8me2s, H4R3me2s, and H3R2me2a in the schizonts of PfPRMT5 disruptant (ΔPfPRMT5-1). H3 was used as a loading control. f Western blots indicate the pattern of overall protein arginine methylation (MMA, aDMA, and sDMA) between ΔPfPRMT5-1 and WT (3D7) parasites. A considerable reduction in the sDMA level was detected in ΔPfPRMT5-1. PfHSP70 was used as an equal loading control. The relative levels of MMA, aDMA, and sDMA were normalized with PfHSP70. The standard deviation was shown as error bars.

Article Snippet: Western blots were performed with anti-MMA, -aDMA, and -sDMA antibodies (Cell Signaling Technology, Inc.) as the primary antibodies.

Techniques: Disruption, Control, Western Blot, Methylation, Standard Deviation