SHP-2 is downstream of Fer in mediating β-catenin regulation during presynaptic development. (A–F) Effects of PTP1B and SHP-2 depletion on synaptic vesicle localization and suppression of SHP-2 phenotype by overexpression of β-catenin (β-cat) and β-catenin Y654F. 14 DIV hippocampal neurons were imaged at 14 DIV after transfection at 10 DIV with synaptophysin-GFP (Syn-GFP) together with the indicated constructs. Quantification of the synaptophysin-GFP coverage (normalized micrometers of GFP per 10 μm of axon length) is presented in D–F. SHP-2 shRNA1 was used for all experiments in B, E, and G. In D, n = 22 (vector), 26 (PTP1B shRNA1), 6 (PTP1B shRNA2), 10 (SHP-2 shRNA1), and 6 (SHP-2 shRNA2). In E, n = 12 (vector), 13 (SHP-2 shRNA), 11 (SHP-2 shRNA + β-catenin), and 13 (SHP-2 shRNA + Y654F). In F, n = 11 (vector), 11 (Fer shRNA), and 10 (Fer shRNA + E76K). (G) Increased phosphorylation of β-catenin in SHP-2–depleted neurons. Rat hippocampal neurons were transfected by electroporation before plating and at 14 DIV were lysed and immunoprecipitated with anti–β-catenin. Blots were probed with antiphosphotyrosine mAb 4G10 (left) followed by reprobing with anti–β-catenin. IB, immunoblotting. *, P < 0.05; **, P < 0.01. Error bars represent SEM. Bars, 10 μm.