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Fig. 4 | Cell & Bioscience

Fig. 4

From: Discovery and mechanism studies of a novel ATG4B inhibitor Ebselen by drug repurposing and its anti-colorectal cancer effects in mice

Fig. 4

Ebselen can promote ATG4B oligomerization to highly inhibit ATG4B. A Recombinant ATG4B (5 μM) were incubated with FMK-9a (50 μM) and different concentrations (5, 25 and 50 μM) of Ebselen incubated for different time points (0.5, 1 and 3 h), and the results were detected by non-reducing electrophoresis. B Ebselen (50 μM) was incubated with different concentrations (0.05, 1, 2, 10, and 20 mM) of DTT for 30 min in advance, then recombinant ATG4B (5 μM) was added for another 30 min until submitted to non-reducing electrophoresis. C Recombinant ATG4B (5 μM) was incubated with Ebselen (50 μM) for 30 min before different concentrations (0.05, 1, 2, 10, and 20 mM) of DTT were added until subjected to non-reducing electrophoresis 30 min later. D Recombinant ATG4B WT and 2CS (C292/361S) mutant (5 μM) were treated with or without Ebselen (50 μM) and subjected to reducing or non-reducing electrophoresis. E Mass spectrometry under denaturing condition of ATG4B 2CS mutant treated with Ebselen. F Ebselen (0.5 μM) was incubated with ATG4B WT and 2CS mutant (0.75 μg/ml) for different time (15, 30, 60 and 90 min), and then the inhibition rates were detected by FRET assay

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