Figure 3

Suppression of NEAP expression increases NGF-induced TrkA and FGFR1 phosphorylation. PC12-pSuper and PC12-shNEAP cells were treated with NGF (50 ng/ml) and the cells were collected at the indicated time points. (a) Levels of TrkA, pTrkA, FGFR1 and pFGFR1, and NEAP were examined by immune-blotting assays using specific antibodies as indicated. The pTrkA/TrkA and pFGFR1/FGFR1 ratios in the untreated PC12-pSuper cells were designed as 1. The results from two independent experiments were presented in Supplementary Materials. (b) The cells were pre-treated with puromycin (5 μg/ml) for 2 h and then stimulated with or without NGF (50 ng/ml) for the indicated time periods. The levels of TrkA, NEAP, and β-actin were examined by immune-blotting assays. The ratios of TrkA/actin were determined for each sample and its values in untreated PC12-pSuper and shNEAP cells were designed as 1. The results (mean ± standard deviation) from three immuno blots were analyzed by the Student’s t test. The time points with significantly lower TrkA expression (p < 0.05) in comparison with the untreated control were indicated with asterisks.