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. 2023 Jul 3;18(7):1594–1610. doi: 10.1021/acschembio.3c00212

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Inhibitor transformation in cell lysates. Panel (A): Western blot analysis of HDAC6 expression in parent HEK293T cells (HEK) and HEK293/HDAC6 (HDAC6) stable transfectants. A-tubulin was used as a loading control. The asterisk and arrow mark the HALO-HDAC6 fusion and endogenous HDAC6, respectively. Panels (B, C): Kinetics of the hydrolysis of 8 in lysates of HEK293T (B) and HEK293/HDAC6 (C) cells. Compound 8 (20 μM final concentration) was mixed with cell lysates (final protein concentration of 2.5 mg mL^–1), and compound hydrolysis at 37 °C was monitored by LC–MS/MS upon inhibitor extraction by acetonitrile precipitation.