Figure 5

Expressions by RT-PCR and western blot in bladder cell lines. (A) Netrin-1 & UNC5B mRNA expression in four bladder cell lines (T24, SV, 5637, and BIU-87) was assayed using quantitative real-time PCR. To normalize Netrin-1 & UNC5B expression, β-actin was used as internal control. Each expression level was shown as mean + SD. UNC5B expression was significantly greater in SV cells than in 5637, T24 or BIU-87, while netrin-1 was lower in SV than in BC cell lines. *P < 0.05 compared with BC cell lines including 5637, T24 and BIU-87 (paired t-test). (B) UNC5B &(D) Netrin-1 expression at protein level in different cell lines (SV, BIU, 5637 and T24) were analyzed by western blots with 50 μg protein extracts. GAPDH was selected to be endogenous control. The differences in UNC5B expression between 5637, T24 and BIU-87 were significant. SV cells represent bladder normal cell lines of which UNC5B expression is the most, the others are BC cell lines, and netrin-1 expresses the least in SV. (C) & (E) The ratio between the optical density of UNC5B & netrin-1 and GAPDH of the same tissue was calculated and expressed graphically (P < 0.05). *P < 0.05 compared with BC cell lines (paired t-test).