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. 2014 May 15;4:4972. doi: 10.1038/srep04972

Figure 5. NgR1-phosphorylation is essential for enhancement of neuronal cell production induced by treatment with PKA and ATP.

Figure 5

Plasmids encoding either wild type NgR1 or S281A mutant NgR1 were transfected into NSPs before the induction of differentiation. Expression of wild type and mutant NgR1 were examined by Western blotting (a) and immunofluorescence analysis (b). In (a), 40 µg of each protein sample was analysed by SDS-PAGE. Actin was employed as an internal standard. The bars indicate 25 µm. (c) At the Day 10 of differentiation with or without PKA plus ATP, cells were fixed and stained for GFAP, beta III tubulin and DAPI. The bars indicate 25 µm. In (d), cells transfected with either wild type or the S281A mutant NgR1 were treated as in (c) and manually counted. Each experiment was repeated four times, and each bar and error bar represents the average proportion and the S.D., respectively. Differences between the proportions of beta III tubulin-positive cells in the cells transected with mutant NgR1 were not statistically significant. (*p < 0.05, Student's t-test).