Ndr2-T442D is sufficient to rescue the suppression of Rassf5. (A) Schematic representation of Ndr2. Mst kinases phosphorylate Ndr2 at Thr442 (p) and activate its kinase activity. (B) Hippocampal neurons were transfected at 0 DIV with vectors for GFP (green), phospho-mimic Ndr2-Thr442 (N2-442D), non-phosphorylatable Ndr2-Thr442 (N2-442A) and an shRNA against Rassf5 or pSM2 (control), as indicated. Neurons were fixed at 3 DIV and stained with the Tau-1 antibody (blue) and an anti-MAP2 antibody (red). Scale bars: 50 µm. (C) The percentage of unpolarized neurons without an axon (0, white), polarized neurons with a single axon (1, gray) and neurons with multiple axons (>1, black) is shown. Values are the mean±s.e.m. (three transfections, n>150 neurons); ns, non-significant; *P<0.05; ***P≤0.001, compared with control (GFP+pSM2); two-way ANOVA.