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. Author manuscript; available in PMC: 2015 Apr 23.
Published in final edited form as: Mol Cancer Res. 2014 Feb 10;12(4):527–538. doi: 10.1158/1541-7786.MCR-13-0567

Figure 6. Regulation of TXNDC5 transcriptional activity by NR4A1.

Figure 6

(A) Panc-1 cell swere transfected with siNR4A1 for 72 hr (left panel) or treated with DIM-C-pPhOH for 24 hr (middle and right panel), and TXNDC5 protein and mRNA levels were determined by western blot analysis and real-time PCR, respectively. (B, left panel) A putative NBRE in the TXNDC5 promoter. (B, middle panel) Cells were cotransfected with each siRNA and pTXVDC5-Luc (−1444/+25), and luciferase activity (relative to β-galactosidase activity) was determined. (B, right panel) Panc-1 cells were transfected with pTXVDC5-Luc (−1444/+25) for 4 hr and treated with DIM-C-pPhOH for another 18 hr. Luciferase activity (relative to β-galactosidase activity) was determined, and the corresponding empty vector was used as a control. (C, left panel) ChIP assay. Panc-1 cells were treated with DIM-C-pPhOH for 6 hr, and the ChIP assay was performed as described in the Materials and Methods. (C, right panel) DNA binding assay. Nuclear extracts of Panc-1 cells were tested for NR4A1-DNA binding activity as described in Materials and Methods. (D) Schematic diagram illustrating induction of apoptosis by inactivation of NR4A1.