Fig. 3

Netrin-1 induced the anti-apoptotic effect of REH cells through the Unc5b receptor. A Real-time PCR analysis of the expression of netrin-1 receptor in REH cells. ACTIN was used as an internal control. B An anti-His-tag antibody was used to pull down the histagged netrin-1 protein after exogenous recombinant netrin-1 treatment, followed by immunoblotting analysis of the receptor and netrin-1 levels in the precipitation. C The expression of Unc5b was efficiently decreased following transfection with UNC5B interference lentivirus in REH cells. D The 450 nm absorbance of the 3 day growth curve of shCtrl cells, shCtrl cells treated with netrin-1(100 ng/ml), shUNC5B cells and shUNC5B cells treated with netrin-1(n = 3, **P < 0.01, ***P < 0.001, ****P < 0.0001). E The expression levels of CDK4, PCNA, Bcl-2 and Bax in shCtrl cells, shCtrl cells treated with netrin-1 alone (100 ng/ml, 24 h), shUNC5B cells and shUNC5B cells treated with netrin-1. The expression level was detected by western blotting. The expression of Gapdh was applied as an internal control. F Flow cytometric analysis of the apoptotic ratio of shCtrl cells, shCtrl cells treated with netrin-1 alone (100 ng/ml, 24 h), shUNC5B cells and shUNC5B cells treated with netrin-1. G Column graph of the early apoptotic ratio of shCtrl cells, shCtrl cells treated with netrin-1 alone, shUNC5B cells and shUNC5B cells treated with netrin-1. (n = 3, *P < 0.05, **P < 0.01)