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Fig. 1. Expression of complexin II in RBL-2H3 cells. (A) RT-PCR products amplified with specific primer pairs for complexins I and II were electrophoresed in agarose gel (1.5%). Far left and far right lanes are for the 100 bp ladder marker (M). PCR products amplified with primer pairs of complexin I (CPX I) and II (CPX II) using templates derived from rat brain (positive control) and RBL-2H3 cells are shown. The expected size of the product is about 400 bp for both isoforms. A single band in the lane for complexin II of RBL-2H3 cells was detected, and the product was identified as complexin II by DNA sequencing. (B) Western blot analysis for complexin. Cell lysates were prepared from RBL-2H3 cells, P815 cells and rat cerebrum and were electrophoresed by SDS-PAGE. After the samples were transferred to a PVDF membrane, blots were probed with primary antigens specific for complexins I and II. Blots were visualized with anti-mouse IgG conjugated with horseradish peroxidase using chemiluminescence methods. Complexin II was detected at about 19 kD (lower panel), but complexin I was not detected (upper panel) in mast cells. (C) Intracellular distribution of complexin II. Complexin II was detected with anti-complexin II antibody and visualized with FITC-conjugated anti mouse IgG. Fluorescent images were collected with a confocal laser scanning microscope. Complexin II was detected in the cytoplasm and the nucleus.
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