Fig. 5

Inhibition of FAK and Erk 1/2 could reduce REH cells survival. A The 450 nm absorbance of the 3 day growth curve of REH cells (ctrl), REH cells treated with netrin-1 alone(100 ng/ml), REH cells treated with PF-573228 alone (20 nM) and REH cells co-treated with netrin-1 and PF-573228(n = 3, ***P < 0.001, ****P < 0.0001). B The 450 nm absorbance of the 3 day growth curve of REH cells (ctrl), REH cells treated with netrin-1 alone (100 ng/ml), REH cells treated with Magnolin alone (87 nM) and REH cells co-treated with netrin-1and Magnolin (n = 3, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001). C The total protein expression level and phosphorylation level of FAK, c-Raf, Mek1/2 and Erk1/2 in REH cells treated with netrin-1 alone (100 ng/ml, 24 h) and REH cells co-treated with netrin-1 and PF-573228(20 nM, 24 h). The expression level of Gapdh was applied as an internal control. D The expression levels of CDK4, PCNA, Bcl-2 and Bax in REH cells (ctrl), REH cells treated with netrin-1 alone (100 ng/ml, 24 h), REH cells treated with PF-573228 alone (20 nM, 24 h) and REH cells co-treated with netrin-1 and PF-573228 were detected by western blotting. The expression level of Gapdh was applied as an internal control. E The expression level of CDK4, PCNA, Bcl-2 and Bax in REH cells (ctrl), REH cells treated with netrin-1 alone (100 ng/ml, 24 h), REH cells treated with Magnolin alone (87 nM, 24 h) and REH cells co-treated with netrin-1 and Magnolin were detected by western blotting. The expression level of Gapdh was applied as an internal control. F Flow cytometric analysis of apoptotic ratio of REH cells (ctrl), REH cells treated with netrin-1 alone (100 ng/ml, 24 h), REH cells treated with PF-573228 alone (20 nM, 24 h) and REH cells co-treated with netrin-1 and PF-573228. G Column graph of the early apoptotic ratio of REH cells (ctrl), REH cells treated with netrin-1 alone, REH cells treated with PF-573228 alone and REH cells co-treated with netrin-1 and PF-573228 (n = 3, *P < 0.05, **P < 0.01). H Flow cytometric analysis of apoptotic ratio of REH cells (ctrl), REH cells treated with netrin-1 alone (100 ng/ml, 24 h), REH cells treated with Magnolin (87 nM, 24 h) and REH cells co-treated with netrin-1 and Magnolin. I Column graph of the early apoptotic ratio of REH cells (ctrl), REH cells treated with netrin-1 alone, REH cells treated with Magnolin alone and REH cells co-treated with netrin-1 and Magnolin (n = 3, *P < 0.05, ****P < 0.0001).