Figure 9. FEZ1 is involved in Netrin-1 and Sema3A-induced dendritic development. A, B, Images of FEZ1 sgRNA or CRMP1 sgRNA, LUC sgRNA, and uninfected neurons treated with Netrin-1 or Sema3A. Neurons were fixed and stained for MAP2, Tau, and FEZ1, or CRMP1 24 h post-treatment. Scale bars: 20 μm. C, D, Sholl analyses and AUC of FEZ1-deficient and control neurons treated with Netrin-1 or Sema3A. Dendritic complexity of control neurons but not FEZ1-deficient neurons were observed to increase after Netrin-1 or Sema3A treatment. FEZ1 sgRNA control n = 32, netrin n = 35, sema n = 33; LUC sgRNA control n = 31, netrin n = 32, sema n = 35; uninfected control n = 36, netrin n = 38, sema n = 36 collected over three independent experiments. E–G, Total axon length, longest contiguous axon, and axonal branching quantifications for FEZ1-deficient and control neurons treated with Netrin-1 and Sema3A. FEZ1-deficient neurons did not respond to either guidance cues. FEZ1 sgRNA control n = 31, netrin n = 30, sema n = 30; LUC sgRNA control n = 30, netrin n = 30, sema n = 30; uninfected control n = 30, netrin n = 30, sema n = 31 collected over more than three independent experiments. H, I, Sholl analyses and AUC of CRMP1-deficient and control neurons treated with Netrin-1 and Sema3A. CRMP1-deficient neurons displayed an increase in dendritic complexity in response to Netrin-1 but not to Sema3A. CRMP1 sgRNA control n = 37, netrin n = 33, sema n = 35; LUC sgRNA control n = 37, netrin n = 33, sema n = 33; uninfected control n = 39, netrin n = 31, sema n = 32 collected over three independent experiments. J–L, Total axon length, longest contiguous axon and axonal branching quantifications for CRMP1-deficient and control neurons treated with Netrin-1 and Sema3A. No significant differences between treated and untreated neurons were observed for all variables measured in the control groups. CRMP1-deficient neurons displayed increased total axonal length and longest contiguous axon length when treated with Netrin-1 but not Sema3A. CRMP1 sgRNA control n = 30, netrin n = 31, sema n = 30; LUC sgRNA control n = 31, netrin n = 31, sema n = 30; uninfected control n = 31, netrin n = 31, sema n = 31 collected over more than three independent experiments. For all analyses, statistical significance was determined using Kruskal–Wallis analysis. All error bars represent SEM.