Figure 5
Figure 5. C17 causes the accumulation of HMW-Ub and reduce the levels of Bcr-Abl in K562 cells. A. The accumulation of K48-linked Ub in K562
leukemia cells treated with diphenyl carbonates (50 µM) for 2 h. B. As in A, but Bcr-Abl was measured by immunoblotting with anti-Abl antibodies. C. Dose
dependence of the C17-induced depletion of Bcr-Abl. Intervening lane removed for clarity. D. Quantification of blots as in C. E. K562 cells were treated with
C17 (50 µM) in the presence and absence of bortezomib (6 µM) for 4 h and Bcr-Abl was measured by immunoblotting with anti-Abl antibodies. F. Quantitation
of blot in E. Significance: DMSO relative to C17 and bortezomib p=0.002; DMSO relative to C17, p = 0.03. G. Effect of C17 on the cell cycle as determined by
FACS. Black bars represent apoptotic cells (sub G1) and grey bars show cells in G1.
Figure 6
Figure 6. C17 reduces the levels of Mdm2 and cause the accumulation of P53 and P21 in MCF7 cells. Intervening lanes were removed for clarity. A. MCF7
cells were treated with C17 (50 µM) for 4 h and K48 ubiquitin chains were measured by immunoblotting. B-F. MCF7 cells were treated with C17 for 4 h and
the levels of Mdm2 (B), p53 (C, D and E) and p21 (F) were measured by immunoblotting. Note: error bars in E are too small to be clearly observable. G. MCF7
and B16/F10 cells were treated with C17 every 24 h. After 72 h, viable cells were measured by Alamar Blue®. H. The effects of C17 on the cell cycle as
measured by FACS. I. The viability of MCF7 cells treated with products of C17 hydrolysis.
18. Lamberto I, Liu X, Seo HS, et al. Structure-guided development of
a potent and selective non-covalent active-site inhibitor of USP7.
Cell Chem Biol. 2017;24: 1490-1500.
Acknowledgments
MJCL acknowledges a Howard Hughes Medical Institute
International Student Fellowship and a Brandeis Sprout grant.
This work was supported by the National Institutes of Health R01
GM100921 to LH. FEO is a co-founder and shareholder of UbiQ
Bio BV.
19. Turnbull AP, Ioannidis S, Krajewski WW, et al. Molecular basis
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