Fig. 5

Downregulation of ceramide transporter CERT and upregulation of transport protein, ARV-1 in MIA PaCa-2, BxPC3, Panc 1 and HPDE-E6E7 cells treated with GT3. Pancreatic cancer cells MIA PaCa-2, BxPC3, and Panc 1, as well as HPDE-E6E7cells were treated for 4 hours with increasing concentrations of GT3 over a dose range from 0-40μM and probed for CERT and its activated form (a and b for MIA PaCa-2 assays, and g for BxPC3, Panc1, and HPDE-E6E7), ARV-1 (c and d for MIA PaCa-2 assays and f for BxPC3, Panc1, and HPDE-E6E7), and analyzed by western immunoblot. The membranes were reprobed with actin as a loading control. Quantitative RT-PCR using ARV-1 primers was performed on MIA PaCa-2 cells treated with increasing concentrations of GT3 in the same manner as described above (e). All assays were conducted in triplicate and blots shown are representative of the results obtained. For quantification, (graphs) band densities from the treatment conditions identified by the lane labels, were calculated as percentages of the value for the treated, untreated cells (100%). Data shown of representative experiments, (mean ± SE, n = 3). * p <0.05, significant difference between control and GT3 treated cells at different concentrations