1. Activation of CHOP and DR5 by DIM-C-pPhtBu.
Activation of CHOP and DR5 by DIM-C-pPhtBu. A, induction of CHOP and DR5 proteins by DIM-C-pPhtBu. SKOV3 cells were treated with DMSO or 5 to 15 μmol/L DIM-C-pPhtBu for 24 h, and whole-cell lysates were analyzed for CHOP, DR5, and β-tubulin (loading control) by Western blot analysis as described in Materials and Methods. B, activation of pCHOP. SKOV3 cells were transfected with pCHOP, treated with DMSO, 5 to 15 μmol/L DIM-C-pPhtBu, 0.5 μg/mL tunicamycin, and 15 μmol/L rosiglitazone, and luciferase activity was determined as described in Materials and Methods. *, P < 0.05, significant induction, Columns, mean of three replicate determinations for each treatment group; bars, SE. C, activation of pDR5 constructs. SKOV3 cells were transfected with pDR5 constructs treated with DMSO or 15 μmol/L DIM-C-pPhtBu and luciferase activity was determined as described in Materials and Methods. *, P < 0.05, significant induction. D, electrophoretic mobility shift assay analysis. Nuclear extracts from SKOV3 cells treated with DMSO or various ER stress reagents were incubated with −276DR5[32P] and antibodies or oligonucleotides and analyzed in a gel mobility shift assay as described in Materials and Methods.
Ping Lei et al. Mol Cancer Ther 2006;5:
©2006 by American Association for Cancer Research