SrGAP1 and srGAP2 inhibit neuronal differentiation and neurite outgrowth of VPA-induced Neuro2a cells.

<p>A and F. The proper expression of srGAP1-WT/srGAP1^R542A and srGAP2-WT/srGAP2^R527A was confirmed by Western blot with GFP antibody. β-actin was selected as a loading control. B and G. Analysis of cell percentage of VPA-induced Neuro2a cells sharing different length of neurites. <i>n</i> = 3, Mean ± S.D., one-way ANOVA, **<i>P</i><0.01. C–E and H–J. Assessment of neurite outgrowth of Neuro2a cells by VPA stimulation for 24 h. Three endpoints were quantified: total neurite length (C and H), longest neurite length (D and I), and neurite number per cell (E and J). C. Total neurites length of srGAP1 or srGAP1^R542A over-expression (srGAP1-WT, 175.40±17.99 µm; srGAP1^R542A, 156.10±13.84 µm; mock, 162.90±7.87 µm). D. Longest neurite length of srGAP1 or srGAP1^R542A over-expression (srGAP1-WT, 66.98±4.52 µm; srGAP1^R542A, 61.10±2.17 µm; mock, 84.25±6.88 µm). E. Neurite number of srGAP1 or srGAP1^R542A over-expression (srGAP1-WT, 2.128±0.059; srGAP1^R542A, 2.114±0.056; mock, 2.028±0.071). H. Total neurites length of srGAP2 or srGAP2^R527A over-expression (srGAP2-WT, 159.00±11.20 µm; srGAP2^R527A, 138.90±3.27 µm; mock, 143.70±2.11 µm). I. Longest neurite length of srGAP2 or srGAP2^R527A over-expression (srGAP2-WT, 56.67±1.45 µm; srGAP2^R527A, 55.73±2.44 µm; mock, 84.25±6.88 µm). J. Neurite number of srGAP2 or srGAP2^R527A over-expression (srGAP2-WT, 2.120±0.086; srGAP2^R527A, 2.122±0.048; mock, 2.028±0.071). <i>n</i> = 3, Mean ± S.D., one-way ANOVA, *<i>P</i><0.05; **<i>P</i><0.01.