Fig. 3

Ebselen can covalently bind to ATG4B. A, B Covalently bound molecular docking results and schematic sketch of ATG4B (PBD ID: 2Z0E) and Ebselen. Docking results plotted using PyMol. Pink as protein ATG4B, and blue as Ebselen. C, D Target engagements of Ebselen to recombinant ATG4B and C74S mutant protein in vitro thermal shift assay as indicated. E, F Mass spectrometry under denaturing condition of ATG4B only. G Sample from the mass spectrometry assay was subjected to western blot and validated with antibody against ATG4B. H, I Mass spectrometry under denaturing condition of ATG4B treated with Ebselen. J Samples from the mass spectrometry assay were subjected to reducing and non-reducing SDS-PAGE. S.E. as short exposure, L.E. as long exposure