Figure 1

Expression profile of MUC4 in MUC4-transfected SKOV3 cells. (A) Western blot analysis of MUC4 expression in the vector-transfected (SKOV3-Vec) and MUC4-transfected (SKOV3-MUC4) SKOV3 cell line. The MUC4-expressing pancreatic cancer cell line CD18/HPAF was used as a positive control. A total of 20 μg protein from cell extracts was resolved by electrophoresis on a 2% SDS-agarose gel, transferred to the PVDF membrane and probed with anti-MUC4 mouse monoclonal antibody (8G7). The membrane was then incubated with horseradish peroxidase-labeled goat anti-mouse immunoglobulin. The signal was detected using an electrochemiluminescence reagent kit. The MUC4 mucin is a high molecular weight glycoprotein and the predicted size of the transfected MUC4 protein is 320 kDa. (B) Localisation of MUC4 by confocal microscopy. Cells were grown at low density on sterilised cover slips, washed and fixed in ice-cold methanol at −20 °C. After blocking with 10% goat serum, cells were incubated with the anti-MUC4 mouse monoclonal antibody, washed and followed by incubation with FITC-conjugated goat anti-mouse IgG (× 630 original magnification).