Journal: Communications Chemistry

Article Title: A microscale thermophoresis-based enzymatic RNA methyltransferase assay enables the discovery of DNMT2 inhibitors

doi: 10.1038/s42004-025-01439-9

Figure Lengend Snippet: A MST traces of the DNMT2 enzyme reaction (250 nM DNMT2, 5 µM tRNA, 0.9 µM SAM) incubated for variable durations (0–120 min) and yielded significant thermophoresis shifts. B Substrate conversion plots (F norm vs incubation time) reveal steady-state DNMT2 kinetics. C Dose-response curves of aptaSAH1&2 (both 50 nM) in the presence of SHO108/SH112 inhibitors (0–100 µM) indicated these SAH-analogs are not binding to the aptamer at relevant concentrations. D – F Inhibitor characterization of SHO108 from left to right: MST traces at variable inhibitor concentrations; MST-derived dose-response curves; orthogonal 3 H-incorporation assay-derived dose-response curves from detected counts per minute (CPMA). G – I Inhibitor characterization of SH112 analogous to subplot ( D – F ). All inhibitor characterizations were performed in triplicates (mean ± SD, n = 3).

Article Snippet: A compound library for the DNMT2 pilot screening (including Cpd 1–10 ) was commercially obtained (Tocriscreen Epigenetics 3.0, Cat. No. 7578).

Techniques: Incubation, Binding Assay, Derivative Assay

Journal: Communications Chemistry

Article Title: A microscale thermophoresis-based enzymatic RNA methyltransferase assay enables the discovery of DNMT2 inhibitors

doi: 10.1038/s42004-025-01439-9

Figure Lengend Snippet: A Screening a library including 160 drug-like compounds resulted in the identification of ten potential hit compounds (green) with an F norm > 1000‰ (equals >80% inhibition). Initial hits were confirmed by triplicate validation (mean ± SD, n = 3). Positive reaction control: aptamer spiked with 1 µM SAH; negative control: aptamer mock-treated with DMSO. B Exemplary MST traces of the library’s first 80 compounds. C Orthogonal screening of the library by FP displacement experiments using FTAD as the fluorescent tracer. FP experiments reveal that only alexidine ( Cpd 3 , green) is binding to the SAH-binding site. D An inhibition selectivity panel was established by MST aptamer assays using the MTase assays described in this study and 500 µM of the respective hit compound. Only DNMT2 was inhibited significantly. E Alexidine’s ( Cpd 3 ) dose-response curves and K D determination by DNMT2 FP assays (mean ± SD, n = 3). F FP assays to confirm the reversibility of alexidine binding to DNMT2. The polarization of the DNMT2-FTAD complex (96 mP) is displaced in the presence of alexidine (5 mP) and can be effectively restored by analytical size-exclusion chromatography (92 mP). A second treatment with alexidine leads to a repeated FP displacement (4 mP). G Chemical structures of the identified DNMT2 hits.

Article Snippet: A compound library for the DNMT2 pilot screening (including Cpd 1–10 ) was commercially obtained (Tocriscreen Epigenetics 3.0, Cat. No. 7578).

Techniques: Inhibition, Biomarker Discovery, Control, Negative Control, Binding Assay, Size-exclusion Chromatography

Journal: Communications Chemistry

Article Title: A microscale thermophoresis-based enzymatic RNA methyltransferase assay enables the discovery of DNMT2 inhibitors

doi: 10.1038/s42004-025-01439-9

Figure Lengend Snippet: A Screening a library including 80 cysteine-focused covalent compounds resulted in the identification of one single hit compound (green) with an F norm > 1000‰ (equals >90% inhibition). Initial hits were confirmed by triplicate validation (mean ± SD, n = 3). Positive reaction control: aptamer spiked with 1 µM SAH; negative control: aptamer mock treated with DMSO. B Orthogonal screening of the library by fluorescence polarization displacement experiments using FTAD as the fluorescent tracer. FP experiments reveal that only adamantanyl-acryloylurea ( Cpd 11 , green) is binding to the SAH-binding site. C MST-derived dose-response curve for adamantanyl-acryloylurea ( Cpd 11 ) including the chemical structure. All inhibitor characterizations were performed in triplicates (mean ± SD, n = 3). D FP assay with Cpd 11 showing time-dependent DNMT2 inhibition with hyperbolic FP displacement plots. E Covalent dose-response analysis of subfigure D: k obs vs. [I] for the determination of covalent inhibition constants ( K I , k inact ) , .

Article Snippet: A compound library for the DNMT2 pilot screening (including Cpd 1–10 ) was commercially obtained (Tocriscreen Epigenetics 3.0, Cat. No. 7578).

Techniques: Inhibition, Biomarker Discovery, Control, Negative Control, Fluorescence, Binding Assay, Derivative Assay, FP Assay

Journal: International Journal of Biological Sciences

Article Title: ARID1A deficiency activates OSM-STAT3 axis in endometrial cancer, creating vulnerability to JAK/STAT3 inhibition

doi: 10.7150/ijbs.129142

Figure Lengend Snippet: Identification of synthetic lethality of ARID1A and JAK/STAT3 in endometrial cancer through drug library screening. A. Schematic illustration of the synthetic lethality screenings with the Epigenetics Drug Library and the Kinase Inhibitor Library. B. Immunoblot analysis showing ARID1A knockout in A1 and B7 single clone. C. Drugs with selectivity index (SI)≥2 were selected as synthetic lethality candidates. SI=IC50 ARID1A+/+ /IC50 ARID1A-/- . D. Synthetic lethality in HEC1B-ARID1A -/- cell treated with stattic (D, F) and gandotinib (E, G) for 72h. The cell images were taken with IncuCyte ZOOM. Scale bar, 300 μm. H. Immunoblot analysis validated the ARID1A overexpression in HEC1B-ARID1A -/- cell. I. Overexpressed ARID1A rescued the synthetic lethality mediated by stattic. Data are mean±sd. * P <0.05, ** P <0.01. J-K. HEC1B ARID1A isogenic cell pair were transfected with STAT3-siRNA for 72 h. J. The quantitative analysis of synthetic lethality induced by STAT3-siRNA. Data are mean±sd. *** P <0.001. K. The growth curve of HEC1B ARID1A isogenic cell pair treated with STAT3-siRNA. **** P <0.0001. Two-way ANOVA test. L- N. IC50 test treated with stattic and gandotinib in endometrial cancer cell line. Data are mean±sd. * P <0.05.

Article Snippet: Epigenetics Drug Library and the Kinase Inhibitor Library screening were purchased from Selleck Chemicals.

Techniques: Drug discovery, Western Blot, Knock-Out, Over Expression, Transfection

Journal: Scientific Reports

Article Title: Histone acetyltransferase and Polo-like kinase 3 inhibitors prevent rat galactose-induced cataract

doi: 10.1038/s41598-019-56414-x

Figure Lengend Snippet: HAT inhibitor list.

Article Snippet: Epigenetic Multiple Ligand , p300/CBP , Santa Cruz (USA) , 50 μM , × , (29).

Techniques: