Journal: iScience

Article Title: Identification and development of TRPM4 antagonists to counteract neuronal excitotoxicity

doi: 10.1016/j.isci.2024.111425

Figure Lengend Snippet: Identification of five lead TRPM4 antagonist series by high-throughput screening (A) Schematic illustration of the high-throughput screening (HTS) workflow for the identification of TRPM4 antagonists. The HTS involved screening a library comprising 256,286 small molecules. Active compounds were identified using a kinetic FLIPR assay based on membrane-potential sensitive dyes followed by subsequent validation of their activity and determination of IC50 values with an automated patch clamp system (QPatch). (B) Multidimensional scaling (MDS) analysis presenting the distribution of 357 compounds identified during the high-throughput screening. The respective IC50 values determined by QPatch measurements are represented, with compounds exhibiting IC50 values below ≤1 μM highlighted. (C) Assignment of lead compounds and active orthologs to five distinct compound series summarizing chemical features of the identified TRPM4 antagonists. Also see Figures S1–S3 .

Article Snippet: Cells were incubated for 60 min at 37°C before placement into the FLIPR Tetra High-Throughput Cellular Screening System (Molecular Devices).

Techniques: High Throughput Screening Assay, Membrane, Biomarker Discovery, Activity Assay, Patch Clamp