Figure 4.

Genetic Modification of Primary Pancreatic Tumor Cells (PACO2) Using nS/MAR DNA Vector Technology

pS/MAR and nS/MAR vectors expressing the reporter gene GFP were transferred into PACO2 cells through electroporation. (A) The viability and the efficiency (number of viable GFP⁺ cells) of transfection were evaluated by flow cytometry and are represented as boxplots, where the line represents the median (n = 3, analyzed by t test; viability, p = 0.0012; transfection efficiency, p < 0.0001). (B) PACO2 primary pancreatic cancer cells were established with nS/MAR-GFP and n/MAR-SMAD4-GFP (nS/MAR-SMAD4). (C) The expression of the reporter gene GFP was evaluated in flow cytometry in comparison to parental unmodified PACO2 cells. Western blot analysis shows successful re-introduction of SMAD4 in nS/MAR-SMAD4 PACO2 cells. SMAD4 wild-type (WT) Panc-1 cells are shown as a control. (D) Volcano plot showing the gene expression changes of PACO2 GFP versus parental cells. Highlighted in red are genes with a fold change of 2 and p < 0.05.