Figure 2: VLX1570 inhibits and binds to proteasome DUBs in exposed cells. | Scientific Reports

Figure 2: VLX1570 inhibits and binds to proteasome DUBs in exposed cells.

From: The proteasome deubiquitinase inhibitor VLX1570 shows selectivity for ubiquitin-specific protease-14 and induces apoptosis of multiple myeloma cells

Figure 2

(a) Inhibition of active-site-directed labeling of proteasomal deubiquitinases by VLX1570 in MM cells. OPM-2 MM cells were exposed to 0.5 μM VLX1570 for 3 hours and 25 μg whole cell lysates were subsequently labeled with Ub-VS (1 μM), followed by SDS gel electrophoresis and immunoblotting with USP14 or UCHL5 antibodies. The upper bands represent active USP14 or UCHL5 enzymes. (b) Thermo-stabilization of USP14 in exposed cells. The same number of OPM-2 MM cells were exposed to DMSO, 1 μM VLX1570, 1 μM b-AP15, 10 μM b-AP113, 20 μM IU1 or 5 μM WP1130 for 1 hour and analysed for thermostability of UPS14 by CETSA (cellular thermal shift assay)23. The lower panel shows means ± S.E.M. for 3 different experiments (*p < 0.05 by t-test; calculated relative to the inactive control b-AP113). (c) Dose-response of thermal stabilization of USP14. OPM-2 myeloma cells were exposed to different concentrations of VLX1570 for 1 hour and collected for CETSA analysis. Shown are means ± S.E.M. for 3 different experiments (*p < 0.05; **p < 0.01 by t-test).

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