Figure 4. Cortactin is involved in... | Rockefeller University Press

Figure 4.

$Figure 4. Cortactin is involved in shootin1-mediated and L1-CAM–dependent axon outgrowth. (A) Hippocampal neurons overexpressing myc-GST (control) or myc-shootin1 were cultured on coverslips coated with L1-CAM-Fc for 6 d and then immunostained with anti-myc and tau-1 antibodies. The neurons were also cotransfected with a vector to express a control miRNA or cortactin miRNA (#1 or #2), which also coexpresses EGFP. RNAi-refractory cortactin was also coexpressed in the indicated experiments. Arrowheads in the enlargements of the boxed areas of the main images indicate axons labeled by the axon-specific marker tau-1 antibody. The graph shows the percentage of neurons with multiple axons. A total of 509 neurons were examined in three independent experiments. (B) Hippocampal neurons were transfected with a control miRNA or cortactin miRNA (#1 or #2) and cultured on coverslips coated with L1-CAM-Fc (L1-Fc) or polylysine (PDL) for 48 h. RNAi-refractory cortactin was also coexpressed in the indicated experiments. The graphs show axon length. A total of 1,111 neurons were examined in three independent experiments. Data represent means ± SEM (error bars); ***, P < 0.01; **, P < 0.02. Bars, 50 µm.$

Cortactin is involved in shootin1-mediated and L1-CAM–dependent axon outgrowth. (A) Hippocampal neurons overexpressing myc-GST (control) or myc-shootin1 were cultured on coverslips coated with L1-CAM-Fc for 6 d and then immunostained with anti-myc and tau-1 antibodies. The neurons were also cotransfected with a vector to express a control miRNA or cortactin miRNA (#1 or #2), which also coexpresses EGFP. RNAi-refractory cortactin was also coexpressed in the indicated experiments. Arrowheads in the enlargements of the boxed areas of the main images indicate axons labeled by the axon-specific marker tau-1 antibody. The graph shows the percentage of neurons with multiple axons. A total of 509 neurons were examined in three independent experiments. (B) Hippocampal neurons were transfected with a control miRNA or cortactin miRNA (#1 or #2) and cultured on coverslips coated with L1-CAM-Fc (L1-Fc) or polylysine (PDL) for 48 h. RNAi-refractory cortactin was also coexpressed in the indicated experiments. The graphs show axon length. A total of 1,111 neurons were examined in three independent experiments. Data represent means ± SEM (error bars); ***, P < 0.01; **, P < 0.02. Bars, 50 µm.

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