FIGURE 6.

PCDH12 shedding is regulated by various transduction pathways. A and B, CHO-PCDH12 were treated with Me₂SO (NI), 0.1 μg/ml PMA, 10 μm Fsk, 50 μm genistein (Gen), 10 μm 1,2-dipalmitoylphosphatidylinositol 3,4,5-trisphosphate (PIP3), or 10 μm LY294002 (LY). The cell supernatants were harvested at 24 h and analyzed by Western blot, as illustrated in A. M stands for unconditioned cell medium. B, supernatants of three independent experiments were also analyzed by dot blot, and the signal intensities were measured. The data represent the means ± S.D. of signals normalized with signals of controls (NI). Statistical analysis compares controls versus treated cell signals. C and D, CHO-PCDH12 were treated with Me₂SO (NI) or PMA alone (0) or PMA together with either forskolin, genistein, or LY294002. Cell supernatant were harvested and analyzed as above. Statistical analysis compares double treatments to PMA alone. ###, p < 0.001; ##, p < 0.01; #, p < 0.02. E, HUVECs were incubated with Me₂SO (NI), 0.1 μg/ml PMA, 200 ng/ml VEGF, 1 mm histamine (H), 100 nm PGE₂, or VEGF and PGE₂. The supernatants were harvested at 24 h, and 100 μl were used for dot blot analysis. The data are the means ± S.D. of triplicates and are representative of three independent experiments. Significance with NI: ***, p < 0.001; **, p < 0.01; *, p < 0.05. Significance with VEGF alone: Δ, p < 0.01.