**Figure 3.** **Both Nova2 knockdown and dual sevoflurane exposure suppressed Netrin-1/Dcc protein expression.** (**A**) QPCR revealed that Netrin-1 mRNA expression was decreased among 28 candidate genes in the Nova2 shRNA group (*F* = 0.76, *P* = 0.0015**, N = 3, Student’s t-test), while Dcc mRNA expression wasn’t change (F = 0.699, *P* = 0.0519, N = 3, Student’s t-test). (**B**) Western blot analysis demonstrated that Nova2 knockdown suppressed Dcc and Netrin-1 proteins expressions. (**C**) Quantification of the protein expression of Dcc relative to Actin (*P* = 0.0217*, N = 3, Student’s t-test). (**D**) Quantification of the protein expression of Netrin-1 relative to Actin (*P* = 0.0264*, N = 3, Student’s t-test). (**E**) Western blot analysis demonstrated that Nova2 OE upregulated Dcc and Netrin-1 protein expressions in the cortical tissues of offspring mice. (**F**) Quantification of the protein expression of Dcc relative to Actin (*P* = 0.0148*, N = 3, Student’s t-test). (**G**) Quantification of the protein expression of Netrin-1 relative to Actin (*P* = 0.0353*, N = 3, Student’s t-test). (**H**) Western blot analysis demonstrated that dual sevoflurane exposure also decreased Dcc and Netrin-1 proteins expressions. (**I**) Quantification of the protein expressions of Dcc (*F* = 1.118, *P* = 0.0044**, N = 3, Student’s t-test) and Netrin-1 (*F* = 0.386, *P* = 0.0052**, N = 3, Student’s t-test) relative to Actin. **P*<0.05. ***P*<0.01, ****P*<0.001.*****P* < 0.0001.