**Figure 2.** **Nova2 deficiency contributes to the neurotoxicity induced by dual sevoflurane exposure.** (**A**) Western blot analysis demonstrated that dual exposure to sevoflurane decreased Nova2 expression in the cortical tissues of offspring mice. (**B**) Quantification of the protein expressions of Nova2 relative to Actin (*P* = 0.0174*, N = 3, Student’s t-test). (**C**) Nova2 shRNA (sh3) reliably reduces the Nova2 expression. (**D**) Quantification of the protein expressions of Nova2 relative to Actin (*F* = 13.54, *P* = 0.0146*, N = 3, one-way ANOVA). (**E**) Nova2 vector significantly increased the Nova2 expression. (**F**) Quantification of the protein expressions of Nova2 relative to Actin (*P* = 0.0007***, N = 3, Student’s t-test). (**G**) Nova2 knockdown significantly decreased the neuronal migration cortex of offspring mice, while overexpression (OE) of Nova2 reversed loss of function. Scale bars=200 μm. (**H**) Quantification of GFP+ cells at different positions in different groups. Compared to the Con-sh group, the Nova2-sh group had significantly larger fractions of neurons in the IZ (43.86 ± 2.49%, *P* = 0.0021**, N = 3, Student’s t-test) and the VZ/SVZ (14.84 ± 2.4 %, *P* = 0.0283*, N = 3, Student’s t-test), and a significantly smaller fraction of neurons in the UpCP (28.68 ± 2.2768%, *P* = 0.0098**, N = 3, Student’s t-test). (**I**) Quantification of GFP+ cells at different positions in different groups. Nova2 OE reversed dual sevoflurane-induced neuronal migration deficits, neurons primarily migrated out of the IZ (4.735 ± 5.084%, *P* = 0.009**, N = 3, Student’s t-test), and were positioned within the UpCP (80.015 ± 4.1507%, *P* = 0.0034**, N = 3, Student’s t-test) and LoCP (10.47 ± 4.483%, *P* = 0.1957, N = 3, Student’s t-test). (**J**) Nova2 knockdown decreased axon length in primary cultured mouse cortical neurons (*P* = 0.0106*, Student’s t-test), while Nova2 overexpression reversed dual sevoflurane-induced axon growth deficits (*P* = 0.0439*, Student’s t-test). (**K**) The statistical results for the axon length in different groups. Scale bars = 100 μm. **P*<0.05. ***P*<0.01, ****P*<0.001.*****P* < 0.0001.