Figure 1. Analysis of human palladin isoforms in pancreatic tissues.

A. Human palladin isoforms. Proline-rich domains are represented by red boxes, and Ig-like domains are shown as blue boxes. The epitope recognized by the 1E6 and 4D10 antibodies is highlighted in green. The region amplified by RT-qPCR is highlighted in light blue. Isoform #1, 3 and 4 are the primary products of the palladin gene and have been detected by immunoblotting. The sequences of these isoforms are published. The sequences of isoforms #2, 5, 6, and 7 were obtained from genomic databases. “ND”: not-determined. B. Western blot analysis of pancreas samples. Small pieces of fresh tissue were snap-frozen in liquid nitrogen, ground in a chilled mortar and pestle, extracted in a detergent-containing lysis buffer, and centrifuged at 15,000×g to remove any unsolubilized particulates. The supernatant was boiled in Laemmli sample buffer and resolved by SDS-PAGE, with 15 µg protein loaded per lane. The samples were immunoblotted and probed with two anti-palladin antibodies and an antibody to GADPH (a housekeeping gene) as a control for equal loading. Lanes 1–2: normal pancreas. Lanes 3–5: primary adenocarcinoma tumors (PDA). Lane 6–7: Non-primary adenocarcinoma tumors (Non-PDA) (Lane 6: solid pseudopapillary tumor, Lane 7: neuroendocrine tumor). C. RT-qPCR. Total RNA was isolated from normal tissue (patients 1–4) and PDA tumors (patients 1–3), reverse transcribed, and subjected to RT-qPCR using gene-specific primers. Each bar represents the mean + SEM (0.06–0.35%) from three or more independent determinations.