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. 2006 Apr 5;26(14):3829–3839. doi: 10.1523/JNEUROSCI.4247-05.2006

Figure 5.

Figure 5.

Effects of increased NG2 expression on neurite growth. A–D, Comparison of the level of NG2 expression in control NG2 cells and in NG2 cells infected with adeno-NG2. A, B, Surface immunofluorescence labeling (live-cell labeling) for NG2 on uninfected (A) and adeno-NG2-infected cells (B; moi 5). Images were acquired under identical exposure conditions below pixel saturation. Scale bar: (in B) A, B, 12 μm. C, Quantification of fluorescence intensity of uninfected NG2 cells and NG2 cells infected with adeno-NG2. Cells transduced with adeno-NG2 expressed fivefold more surface NG2 than uninfected NG2 cells. D, Western blots comparing the amount of NG2 in extracts of uninfected and adeno-NG2-infected NG2 cells. The same blot was probed with antibodies to NG2 (top) and GAPDH (bottom) chABC+: extracts treated with chondroitinase ABC. Smaller amounts of protein extracts from adeno-NG2-infected cells were loaded to allow for densitometric analyses of the samples within the dynamic ranges of the pixels, as seen by the weaker GAPDH bands in lanes 3 and 4. The positions of molecular weight standards are indicated on the left. E–I, Response of P1 hippocampal neurons to NG2 cells expressing elevated levels of NG2. E, F, Double-immunofluorescence labeling for NG2 (red) and βIII tubulin (green) to illustrate the spatial relationship between neurites and NG2 cells expressing normal (E) or increased levels of surface NG2 after adeno-NG2 transduction (F; moi 5). Scale bar: (in F) E, F, 24 μm. G, Percentage of neurite tips contacting uninfected or adeno-NG2-infected NG2 cells. ns, No statistically significant difference between the number of neurite tips contacting uninfected and adeno-NG2-infected NG2 cells (P = 0.239, t test). H, Total neurite lengths of neurons grown on uninfected or adeno-NG2-infected NG2 cells. ns, No significant difference (P > 0.05, one-way ANOVA). They were significantly longer than neurite lengths on PLL (P < 0.001, one-way ANOVA). I, The lengths of the longest neurite of neurons grown on uninfected or adeno-NG2-iinfected NG2 cells. ns, No significant difference (P > 0.05, one-way ANOVA). They were significantly longer than neurite lengths on PLL (P < 0.001, one-way ANOVA). Error bars indicate SE.

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