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. 2017 Apr 20;8(31):51296–51316. doi: 10.18632/oncotarget.17261

Figure 4. PEITC reduces levels of USP1 by increasing levels of poly-ubiquitinated USP1.

Figure 4

(A) HEK293T cells were incubated with PEITC (15 μM), bortezomib (300 nM), PEITC (15 μM) together with bortezomib (300 nM) or with DMSO (vehicle only) for 3 h at 37°C. WCLs analyzed by SDS-PAGE and western blot. Data represent three independent experiments performed in duplicate. (B) As in (A) Data represent mean ± SD of 3 independent experiments performed in duplicate. Representative blot shown in (A). (C) HEK293T cells were transiently transfected with HA-Ubiquitin using TransIT2020 transfection reagent. 24 h post-transfection, cells were incubated with PEITC (15 μM), bortezomib (300 nM), PEITC (15 μM) together with bortezomib (300 nM) or with DMSO (vehicle only) for 3 h at 37°C. USP1 was immunoprecipitated and eluants were probed with anti-HA antibody and anti-USP1 antibody. Actin is shown as a loading control.