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Fig. 4. Either laminin or NGF alone is sufficient to mediate axon assembly from PCL neurons. (A) PCL neurons were unable to extend axons when cultured on poly-D-lysine (PDL) in the absence of extracellular cues. Neurons plated on laminin-coated coverslips extended axons robustly. (B) Neurons did not extend axons when cultured on poly-D-lysine-coated coverslips in serum-free medium. Addition of NGF induced robust axon assembly from the PCL neurons on poly-D-lysine. Magnification, 4x. (C) Quantification of laminin-induced axon assembly from PCL neurons. Please note that the addition of laminin (S, 20 µg/ml) directly to the culture medium induced a similar extent of axon assembly to that of coated laminin (C). Laminin-induced axon growth was independent of the neurotrophin receptor Trk activity. *P<0.0001 vs control. (D) Laminin-induced axon growth was significantly blocked by an integrin function-blocking antibody (20 µg/ml), whereas control IgG protein had no effect. *P<0.0001 vs control. (E,F) Quantification of NGF-induced axon growth from the PCL neurons. Note that other growth factors, such as EGF and IGF, failed to induce axon growth. In addition, inactivation of Rho kinase with inhibitor Y27632 (25 µM) had minimal effect on inducing axon assembly from the PCL neurons, only slightly enhancing basal axon growth. *P<0.005 and ^#P<0.05 compared with levels in the controls.

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