Figure 4.

Neurite outgrowth in PC12 cells transfected with LanCL1 mutants in response to NGF treatment. (A) Immunoblot analysis of the overexpression of myc-tagged LanCL1 proteins in PC12 cells. (Top) Expression levels of the transfected LanCL1 constructs as probed by the anti-myc antibody. (Bottom) Actin level in each sample as the loading control was detected by anti-actin antibody. The vector lanes contain samples from cells transfected with the empty pBI/eGFP vector. (B) Representative fluorescence microscopy images of the cells after a 4-d treatment with 50 ng/mL NGF. Cells ware transfected either with the empty pBI/eGFP vector or with constructs expressing the indicated LanCL1 variants. Bar, 50 μm. (C) Quantification of PC12 cell differentiation. Upon expression of the indicated LanCL1 variants, the percentage of differentiated cells with extended neurites was determined among the transfected cells expressing eGFP. In each case, 500 transfected cells were measured, and error bars represent SEM of at least three independent experiments.