Fig. 2. Ethanol extract of Chaenomeles sinensis upregulates ARE-dependent NRF2 target gene expression in HaCaT cells. (A) Ethanol extract of Chaenomeles sinensis were treated to HaCaT-ARE-GFP-luciferase cells at various concentrations for 24 hours, and the luciferase activity was measured. Asterisks indicate a statistical significance with ***P < 0.001 (n = 3). (B) Ethanol extract of Chaenomeles sinensis does not affect the viability of HaCaT cells. Ethanol extract of Chaenomeles sinensis was treated to HaCaT cells at various concentrations for 24 hours, and the MTT assay was conducted. (C) Ethanol extract of Chaenomeles sinensis induces NRF2 expression in HaCaT cells. Ethanol extract of Chaenomeles sinensis were treated to HaCaT cells at various concentrations for 24 hours, and Western blot analysis was conducted against NRF2. (D) Ethanol extract of Chaenomeles sinensis activates transcription of NRF2 target genes (HO-1 and NQO1). Ethanol extract of Chaenomeles sinensis (800 μg/mL) was treated to HaCaT cells for 24 hours, and real-time RT-PCR was conducted using HO-1 and NQO1 specific primers. Asterisks indicate a statistical significance with ***P < 0.001 (n = 4). ARE, antioxidant response element; NRF2, NF-E2-related factor 2; HO-1, heme oxygenase-1; NQO1, NAD(P)H:quinone oxidoreductase.

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