Avn C induces heme oxygense-1 expression through increased DNA binding activity of Nrf2 in 2DD fibroblasts. (A) 2DD human skin fibroblasts were treated with DMSO (control) or Avn C (50 μM, 100 μM and 200 μM) for 48 h. qRT-PCR on cDNA libraries was performed for glutathione synthetase (GSS), heme oxygenase 1 (HMOX1), superoxide dismutase 1 (SOD1), glutathione superoxidase 1 (GPx1) and catalase (CAT) gene transcripts. Data are represented as fold changes against control. (B) Western blot analysis of heme oxygenese-1 (HO-1) expression in 2DD cells treated with DMSO (control) or Avn C (100 μM and 200 μM) for 48 h. Data are presented as a ratio of HO-1 to β-actin. Representative blot images are shown. (C) ChIP-qPCR analysis of Nrf2 binding activity to promoter regions of heme oxygenase 1 (HMOX1) and NAD(P)H quinone dehydrogenase 1 (NQO1) in 2DD cells treated with DMSO (control) or 100 μM Avn C for 48 h. Data are represented as the enrichment relative to % input. All graphs indicate the average values from three biological replicates. Error bars represent the standard deviations. *p values < 0.05, **p values < 0.01 vs control.