Figure 3.

Phosphorylated JNK and the dependent invasion/angiogenesis are inversely correlated with MKP-1 expression in human urothelial carcinoma cells. A and B: The human urothelial carcinoma cell lines UMUC-6 and UMUC-14 were transfected with pTK-GFP and after a 24-hour cultivation were treated with or without 25 μmol/L SP600125 or 1 μmol/L JNK inhibitory cell-permeable peptide for 1 hour then they were seeded on top of CAMs and incubated for 3 days. A: Immunohistochemical findings of cross sections using anti-GFP antibody are shown. Invasion was assessed by counting the number of invading cells per high-power field and quantifying front depth of invasion (three or more cells), whereas vessels were enumerated by counting vessel branch points in a double-blinded manner. B: Each value is the mean ± SE from at least three experiments. C: Samples obtained from UMUC6 and UMUC14 cells cultured in vitro or grafted on top of CAMs were lysed with lysis buffer as described in Materials and Methods, and expression of phosphorylated JNK, JNK, and MKP-1 was examined by immunoblotting using anti-phosphorylated JNK, anti-JNK, and anti-MKP-1 antibodies.