Adaphostin (ADA) effect on HMOX1 related genes, ROS, and HMOX1 protein. (A) ADA modulation of NRF2, HMOX1, GCLC, and NQO1 gene expression. Cells were treated with 1 μM of ADA for 1, 6 and 24 h and gene expression was measured by microarray and quantitative RT/PCR and expressed as fold change of drug -treated NRF2, HMOX1, GCLC, and NQO1 compared with control (n = 4; +/- SD). Both HMOX1 and NQO1 were significantly up-regulated by ADA (** p < 0.01). (B) Increased ROS production after ADA treatment. Cells were treated for 2 and 4 h with 1 μM ADA and ROS was measured using DCFH-DA (10 μM). There was a significant increase in ADA-induced ROS production. After 2 and 4 h (n = 2 +/- SD, * p < 0.05). (C) ADA induces HMOX1 protein. NCI-H522 cells were incubated for 2 h, 4 h and 6 h with 1 μM of ADA and whole cell extracts were resolved by Western blot analysis as indicated in the Materials and Methods. Data are representative of three independent experiments.