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. 2022 Jun 30;13:3778. doi: 10.1038/s41467-022-30463-9

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — A Activity of GSK2830371 in the FDP (red) and p38 MAPK (black) enzymatic assays. FDP is cleaved by PPM1D in a substrate-independent fashion while p38 MAPK threonine 180 is dephosphorylated in a substrate-specific manner. N = 3 independent samples runs. Data are presented as mean values ± SD. B Mechanism of inhibition studies using non-linear fit of Michaelis–Menten enzyme kinetics where hydrolysis of FDP is measured in relative fluorescence units (RFU). Data suggest noncompetitive mode of action with α ≥ 1. N = 2 independent samples runs. Data are presented as mean values ± SD. C Differential scanning calorimetry (DSC) and fluorimetry (DSF) showing the melting temperature of PPM1D_1–420 in the absence (black) and presence (red) of GSK2830371. N = 3 independent sample runs. Data are presented as mean values. D Surface plasmon resonance analysis of the interaction of GSK2830371 with PPM1D_1–420. The K_d is calculated from the k_on and k_off measurements. E Structure of GSK2830371 and the GSK2830371 analog 1d used in the isothermal calorimetry experiments. F The thermodynamic profile of the interaction of the GSK2830371 analog (1d) with PPM1D_1–420 as measured by isothermal calorimetry. N = 3 independent sample runs. Source data are provided in the Source Data file.