Issues

Immunofluorescence analysis demonstrating the cytoplasmic localization of galectin-3 in CD18/HPAF pancreatic cancer cells transfected with scramble control vector. The cells were grown at low density on sterile cover slips for 20 hours. After washing with 0.1 M HEPES-containing Hanks buffer, the cells were fixed in ice-cold methanol at −20°C for 2 minutes. After nonspecific blocking with 10% goat serum, cells were incubated with antigalectin-3 antibody in PBS for 90 minutes at room temperature. Cells were washed 3 to 4 times with PBS containing 0.05% Tween-20 (PBS-T) and then incubated with FITC-conjugated anti-rat secondary antibodies for 60 minutes. The cells were counterstained with propidium iodide. Finally, slides were washed, mounted, and observed under a ZEISS confocal laser-scanning microscope. Photographs were digitally captured by using 510-software. For further details, please see Senapati and coworkers on page 267 in this issue.