ACA-28, an ERK MAPK Signaling Modulator, Exerts Anticancer Activity through ROS Induction in Melanoma and Pancreatic Cancer Cells

Figure 3

ACA-28 increases ROS and Nrf2 protein levels in SK-MEL-28. (a) A representative Western blot analyzing the Nrf2 and GAPDH expression in SK-MEL-28 cells treated with ACA-28 (10 µM) or DMSO for 6 hr, with or without pretreatment with NAC (5 mM) for 1 hr. GAPDH was used as an internal control. (b) Relative quantification of Nrf2 protein levels shown in (a). Data are expressed as the mean ± standard deviation (SD) from three independent experiments. (c) Representative histogram of ROS levels in SK-MEL-28 cells assessed by fluorescence intensity of CellROX Deep Red using flow cytometry. Cells were treated with ACA-28 (20 µM) or DMSO for 1 hr, with or without pretreatment with NAC (5 mM) for 45 min. (d) The mean fluorescence intensity (MFI) of the data is shown in (c). Data are presented as the mean ± standard deviation (SD) from three independent experiments (n.s.: not significant, , , Tukey’s multiple comparison test).

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