FIGURE 2.

Cellular signaling pathway of PlGF-induced PAI-1 expression. A and B, HPMVEC were pretreated for 30 min with pharmacological inhibitors of PI3K, LY294002 (10 μm); Erk, PD98059 (10 μm); P38α, SB203580 (1 μm); JNK, SP600125 (0.1 μm); NADPH oxidase, DPI (10 μm); HIF-1α, ascorbate (25 μm), and NF-κB, sulfasalazine (2 mm) followed by treatment with PlGF for 6 h. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. C and D, HPMVEC were transfected with indicated siRNA or scrambled (sc) siRNA constructs followed by PlGF treatment for 6 h. A and C, RPA analysis of the indicated genes. Where indicated, the vertical lines show repositioned gel lanes. Data are representative of three independent experiments. B and D, PAI-1 release by ELISA. The data are representative of three independent experiments. ***, p < 0.001. ns, not significant. E and F, HPMVEC were treated with PlGF either in the presence or in the absence of pharmacological inhibitors for 30 min. E, the cytosolic extracts were prepared and subjected to Western blotting for phospho-JNK, and the same blot was reprobed for total JNK. Ab, antibody. F, the nuclear extracts were analyzed for phospho-c-Jun, and the same blot was reprobed for total c-Jun and c-Fos. Data are representative of three independent experiments.