Fig. 2

Phospho-mutant 6T/S-A NeuroD4 has increased proneural activity relative to WT NeuroD4 (a-g). a Schematic representation of WT NeuroD4 and full phospho-mutant 6T/S-A NeuroD4 protein sequences, indicating the relative positions of the six SP or TP sites that have been mutated to AP sites. b-g Two-cell stage embryos were unilaterally injected with 100 pg of either WT or 6T/S-A NeuroD4 mRNA, and at stage 18, gene expression was assayed by qRT-PCR (b), or by whole mount ISH (d-g) with representative embryo images shown in (c). For qRT-PCR analysis (b) significance is calculated as described in the methods section for phospho-mutant NeuroD4 relative to WT NeuroD4 (blue adjoining lines and stars) and relative to uninjected control embryos (shown with black stars) [n = 5]; (p < 0.05) = *; (p < 0.025) = **; (p < 0.0125) = ***. For ISH analysis, embryos were scored according to the scale described in Additional file 1 for expression of neural-β-tubulin (D [n = 100–115]), p27Xic1 (E [n = 24-33]), xMyt1 (F [n = 26-32]), and xNeuroD1 (G [n = 23-28]). Views in (C [i-iii]) are dorso-ventral (DV); views in (C [iv]) are rostro-caudal (RC) with dorsal surface facing up, trigeminal ganglia indicated by arrows. All images show injected side to the right