Journal: The International journal of developmental biology

Article Title: A screen of kinase inhibitors reveals a potential role of Chk1 in regulating Hydra head regeneration and maintenance.

doi: 10.1387/ijdb.210087yl

Figure Lengend Snippet: Fig. 1. Schematic illustration of the 80 kinase inhibitor screen. Three kinase inhibitors (KIs) were tested per 12-well plate. For each plate, 60 Hydra gastric regions were isolated by removing the head and foot. For each treatment, 15 gastric regions were rinsed with the respective solution (KI #1, KI #2, KI #3, or control) before dividing them into 3 wells (5 gastric regions per well) containing the respective exposure solutions. Control replicates were exposed to 1% DMSO alone and included on every plate.

Article Snippet: The use of kinase inhibitors to identify potential novel regulators of Hydra regeneration To identify novel signaling pathways underlying Hydra regeneration, an unbiased screen of 80 kinase inhibitors (Tocriscreen Kinase Inhibitor Toolbox Cat. No. 3514) was performed.

Techniques: Isolation, Control

Journal: The International journal of developmental biology

Article Title: A screen of kinase inhibitors reveals a potential role of Chk1 in regulating Hydra head regeneration and maintenance.

doi: 10.1387/ijdb.210087yl

Figure Lengend Snippet: Fig. 2. Morphological categorization following exposure to the 80 kinase inhibitors. In total, 4 major morphologies were observed in Hydra following exposure to the 80 kinase inhibitors: (A) complete regeneration, (B) partial regeneration – the presence of shortened tentacles and/or body column, (C) arrested regeneration - no formation of a head and foot, and (D) disintegration - the breakdown of tissues into debris. Scale bar in (A) represents 1000 mm and applies to all panels.

Article Snippet: The use of kinase inhibitors to identify potential novel regulators of Hydra regeneration To identify novel signaling pathways underlying Hydra regeneration, an unbiased screen of 80 kinase inhibitors (Tocriscreen Kinase Inhibitor Toolbox Cat. No. 3514) was performed.

Techniques:

Journal: The International journal of developmental biology

Article Title: A screen of kinase inhibitors reveals a potential role of Chk1 in regulating Hydra head regeneration and maintenance.

doi: 10.1387/ijdb.210087yl

Figure Lengend Snippet: Fig. 3. Drug Exposure Impact of the 80 kinase inhibitors on Hydra regeneration. Kinase inhibitors were grouped by the main signal transduction pathways they are presumed to target. Drug Exposure Impact (DEI) values on regeneration were calculated as the difference in regeneration scores obtained for control and Hydra exposed to 5 mM kinase inhibitor. DEI values for complete regeneration ranged between -1.07 and 1.2; partial regen eration ranged between -4.4 and 0.6; arrested regeneration ranged between -7.67 and -4; and disintegration ranged between -10 and -7.8. Error bars represent ± 2x standard error.

Article Snippet: The use of kinase inhibitors to identify potential novel regulators of Hydra regeneration To identify novel signaling pathways underlying Hydra regeneration, an unbiased screen of 80 kinase inhibitors (Tocriscreen Kinase Inhibitor Toolbox Cat. No. 3514) was performed.

Techniques: Transduction, Control

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: Cell reports

Article Title: Long-Term In Vitro Expansion of Epithelial Stem Cells Enabled by Pharmacological Inhibition of PAK1-ROCK-Myosin II and TGF-β Signaling

doi: 10.1016/j.celrep.2018.09.072

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Tocriscreen Stem Cell Toolbox , Tocris , Cat# 5060.

Techniques: Virus, Selection, Recombinant, Real-time Polymerase Chain Reaction, Sequencing, Paraffin Section, Staining, Enzyme-linked Immunosorbent Assay, In Vivo, Methylated DNA Immunoprecipitation Sequencing, Cell Culture, Software, DNA Methylation Assay