**Figure 4.** **Netrin-1/Dcc deficiency displayed a similar defect in neuronal migration and axon outgrowth as Nova2 deficiency induced by dual sevoflurane exposure.** (**A**) Dcc shRNA (sh3) reliably reduces the Dcc expression. (**B**) Quantification of the protein expressions of Dcc relative to Actin (*F* = 10.57, *P* = 0.0226*, N = 3, one-way ANOVA). (**C**) Netrin-1 shRNA (sh3) reliably reduces the Netrin-1 expression. (**D**) Quantification of the protein expressions of Netrin-1 relative to Actin (*F* = 7.084, *P* = 0.0445*, N = 3, one-way ANOVA. (**E**) Dcc and Netrin-1 knockdown both significantly inhibited neuronal migration cortex of offspring mice. (**F**) Quantification of GFP+ cells at different positions in different groups. Compared to the Con-sh group, the Dcc-sh group had significantly larger fractions of neurons in the IZ (40.23 ± 1.20208%, *P* = 0.0005***, N = 3, Student’s t-test) and the VZ/SVZ (3.93 ± 2.50315%, *P* = 0.13, N = 3, Student’s t-test), and a significantly smaller fraction of neurons in the UpCP (28.335 ± 1.9728%, *P* = 0.0434*, N = 3, Student’s t-test). (**G**) Quantification of GFP+ cells at different positions in different groups. Compared to the Con-sh group, the Netrin-1-sh group had significantly larger fractions of neurons in the IZ (42.665 ± 6.73872%, *P* = 0.0239*, N = 3, Student’s t-test) and the VZ/SVZ (5.335 ± 1.0818%, *P* = 0.8521, N = 3, Student’s t-test), and a significantly smaller fraction of neurons in the UpCP (26.57 ± 2.0223%, *P* = 0.0037**, N = 3, Student’s t-test). (**H**) Both Dcc and Netrin-1 knockdown decreased axon length of neurons in primary cultured mouse cortical neurons. (**I**) The statistical results for the axon length in Dcc-sh RNA group (*P* = 0.0054**, Student’s t-test) and Netrin-1-shRNA group (*P* = 0.0044**, Student’s t-test). Scale bars = 100 μm; approximately 70 cells from three independent experiments were counted during the statistical analysis. **P*<0.05. ***P*<0.01, ****P*<0.001.*****P*< 0.0001.