Retinal lamination observed in WT and 0.1 mM 5 dpf morphant embryos treated with PTU.

<p>Green: Alexa-fluor-488-phalloidin: actin. Blue: TOPRO: nuclei. Red: Zpr-1: double cone photoreceptors. hc: horizontal cells. os: outer segments. is: inner segments si.</p

Ultra structural architecture analysis of inner ear and stereocilia performed on MO1 morphant embryos using fluorescent staining.

<p>A. Phalloidin staining of WT larvae inner ear highlighting the three sensory areas containing stereocilia. B. Phalloidin staining of 0.1: A and B: 20 µm. C and D: 5 µm.</p

Evolutionary analysis of USH2A isoform_a.

<p>Alignment of the exons that correspond to the hypothetic short isoforms of <i>USH2A.</i> Only in Primates and most Eutherians the “KCV end” seems to be conserved.</p

Primers used to amplify and sequence the full-length open reading frame (ORF) of <i>Ol-Ush2a</i>.

<p>Primers used to amplify and sequence the full-length open reading frame (ORF) of <i>Ol-Ush2a</i>.</p

Medaka embryos obtained after MO1 and MO2 injections.

<p><b>A, B, G, H:</b>MO1 C (A, B) and MO C (G, H) embryos at 72 hpf. <b>C, D, I, J:</b> MO1 (C, D) and MO2 (I, J) injected embryos at 72 hpf, showing a <b>mild phenotype</b>. <b>E, F, K, L:</b> MO1 (E, F) and MO2 (K, L) injected embryos at 72 hpf, showing a <b>severe phenotype</b>. Scale bars: 100 µm. Embryos in frontal (A, C, E, G, I, K) and lateral position (B, D, F, H, J, L).</p

Spatial and temporal <i>Ol-Ush2a</i> expression pattern analysis performed by RT-PCR amplification.

<p><b>A</b>: RT-PCR amplification from adult organs. <b>B</b>: RT-PCR amplification from different embryonic developmental stages.</p

Medaka fish Ol-Ush2a predicted protein.

<p>Motif alignment of the human, murine, zebrafish and medaka USH2A proteins. The two lines beneath the murine Ush2a denote the two predicted variants reported (USH2A_iso_a: 170 kDa and USH2A_iso_b: 600 kDa). Symbols representing different motifs are given at the bottom of the diagram: LamGL, LamG-like jellyroll fold domain; EGF-Lam, laminin-type epidermal growth factor-like domain; FN3, fibronectin type 3; LamG, laminin G domain.</p

RT-PCR results from total RNA extracted from control and morphant MO3 embryos at 96 hpf.

<p>C. Control embryos. Expected size: 573 bp MO. Morphant embryos. Expected size: 444 pb.</p