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. 2013 Sep 17;170(3):489–505. doi: 10.1111/bph.12111

Figure 1.

Figure 1

URB597 induces HO-1 and NQO-1 transcripts in breast cancer cell lines. (A) URB597 increases mRNA levels of HO-1 and NQO-1. Furthermore, 50 ng of total RNA of MCF-7 cells or MDA-MB-231 cells treated with vehicle (DMSO) and URB597 (5 μM) for 6 h was subjected to semi-quantitative RT-PCR to determine the levels of HO-1 and NQO-1 mRNA. β-Actin was used as a control for semi-quantitative RT-PCR. (B) URB597 increases protein levels of HO-1 and NQO-1. 25 μg of total protein extract of MCF-7 cells or MDA-MB-231 cells treated with vehicle (DMSO) and URB597 (5 μM) for 16 h was subjected to detect the levels of HO-1 and NQO-1 protein. Equal loading of proteins was demonstrated by anti-actin antibody. The levels of HO-1 and NQO-1 expression were normalized to actin expression and the relative expression was represented graphically. All the figures are representative of three independent experiments. (C) URB597 activates nqo1-ARE-Luc reporter in MCF-7 cells and MDA-MB-231 cells. In 6 well plates, MCF-7 cells or MDA-MB-231 cells were co-transfected with 0.5 μg of nqo1-ARE-Luc reporter and 0.15 μg of pCMV-β-galactosidase plasmids, followed by DMSO (vehicle) and URB597 (5 and 10 μM) treatment for 16 h. URB597-induced luciferase activity was normalized by β-galactosidase activity. The induction of reporter activation was represented as fold increase with respect to the vehicle-treated control. The data represent the mean ± SD of at least three independent experiments performed in triplicate. *P < 0.05 as compared with the control.