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. Author manuscript; available in PMC: 2023 Feb 17.
Published in final edited form as: Mol Cancer Ther. 2021 Jun 22;20(9):1692–1701. doi: 10.1158/1535-7163.MCT-21-0210

Figure 3. MSU38225 enhances the degradation of Nrf2 via the proteasome system.

Figure 3.

A. A549 cells were treated with 5 μM MSU38225, 5 μM MG132, or the combination for 24 hours, and Nrf2 protein expression evaluated by western blotting. B. A549 cells were treated with either DMSO or MSU38225 for 24 hours, and Nrf2 protein was pulled down using agarose beads followed by immunoblotting with anti-ubiquitin antibodies. On the left, the same amount of total protein was loaded; on the right, the same amount of Nrf2 protein was loaded. C. A549 cells were pretreated with DMSO or 5 μM MSU38225 for 4 hours. After washout with PBS, cells were treated with 1 μM cycloheximide for 0–60 minutes. Nrf2 protein expression was detected and quantified using ImageJ (results were normalized to the respective 0 min time point). Wild type, KEAP1 CRISPR knockout (D) and βTrCP CRISPR knockout (E) Jurkat cells were treated with tBHQ +/− MSU38225 for 24 hours and expression of Nrf2, Keap1, NQO1 and βTrCP was detected by western blotting.