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. 2018 Aug 3;293(37):14417–14428. doi: 10.1074/jbc.RA118.003729

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© 2018 Wu et al.

Published under exclusive license by The American Society for Biochemistry and Molecular Biology, Inc.

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Figure 6. — ACh reduces melanin production when AChE-inhibited B16F10 cells. A, cultured B16F10 cells were treated with various concentrations of ACh with or without pretreatment of BW284c51 (BW) (20 μm) and the cell lysates were collected for determination of intracellular melanin content, pCRE-Luc, pMITF-Luc, and pTYR-Luc. Data are normalized and expressed as the percentage of control (sample treated with vehicle). B, cultured cells were treated with various concentrations of ACh with pretreatment of BW284c51 (20 μm), and cell lysates (40 μg) were collected for Western blot analysis. GAPDH was used as loading control. Data are normalized and expressed as the percentage of control. C, cultured B16F10 cells were transfected with ACHE shRNA or cotransfected with AChE and PRiMA cDNA, then treated with BW284c51 (20 μm) and ACh (1 mm). Cell lysates were collected for Ellman assay and luciferase assay. Data are normalized and expressed as the percentage of control (transfected with empty vector). All values are in mean ± S.D., n = 4, each with triplicate samples; *, p < 0.05; **, p < 0.01.