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. 2021 Nov 24;298(1):101448. doi: 10.1016/j.jbc.2021.101448

Figure 7.

Figure 7

USP10-KD does not reduce the amount of Nrf2 mRNA in neuronal cells.A, SH-SY5Y cells were transfected with USP10-siRNA (USP10-2) or control (NT) using Lipofectamine RNAiMAX. Cells were pretreated with 5 nM Bafilomycin A1 30 min before dopamine treatment and further treated with 0.4 mM dopamine or DMSO with or without 5 μM MG-132 for 8 h. Whole cell lysates prepared from transfected cells were characterized by Western blotting using the indicated antibodies. B, SH-SY5Y cells were transfected with USP10-siRNA (siUSP10-2) or control (siNT) using Lipofectamine RNAiMAX. Cells were treated with 0.4 mM dopamine or DMSO for 3 or 6 h. Total RNA was extracted from these cells, and the relative amount of NQO1, GSTM1, p62, and Nrf2 mRNA to β-actin mRNA was measured by real-time RT-PCR. The data were presented as the means ± SD (n = 3). ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001. NS, Not significant; USP10, ubiquitin-specific protease 10.