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[Preprint]. 2023 May 22:2023.05.22.541727. [Version 1] doi: 10.1101/2023.05.22.541727

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Figure 5: — Compound 2 and analogues disrupt the MSN–CD44 PPI. (A) Structures of compound 2 and analogues, with associated solubility data (PBS, 1% DMSO), and activity in both the MSN–CD44 TR-FRET assay and an unrelated PPI (SYK-FCER1G) TR-FRET assay. Solubility was measured by nephelometry. (B) Dose response of compounds in GST pull down assay. Cell lysates were obtained from cells transfected with full length constructs of GST-tagged MSN and Flag-tagged CD44, and were incubated with the compounds. Immunoblotting was performed to detect GST and Flag, and a representative immunoblot from two experiments is shown. (C) Thermal shift assay, showing representative melting temperature (T_m) curves of MSN with compounds. The average change in T_m (av. ΔT_m), compared to DMSO alone, was determined from two experiments (2a: 1.3 °C at 50 μM; 2b: 1.2 °C at 50 μM).