Figure 2: CDDO-Me inhibits hTERT expression in pancreatic cancer cells. A. Effect of CDDO-Me on hTERT gene expression. Panc-1 and Mia PaCa-2 cells were treated with CDDO-Me (0-0.5 μM) for 5 days and total cellular RNA was prepared using TRI-zole reagent. 1 μg of cellular RNA was reverse transcribed using oligo-dt primer and high fidelity reverse transcriptase. 1 μL of cDNA was amplified using hTERT or GAPDH primers. Amplified products were separated on 2% DNA agarose gel. Gels were stained with ethidium bromide and DNA fragments were identified by base pair size. B. Effect of CDDO-Me on hTERT protein. Panc-1 and MiaPaCa-2 cells were treated with CDDOMe as described above and cell lysates were analyzed for hTERT and p-hTERT protein by western blotting. Each experiment was repeated at least three times.