Fig. 2

Fig. 2 Generating a Panx2 global knockout zebrafish line. a) Exon-intron organization of the zebrafish Panx2 gene (ENSDARG00000063019, chromosome 18 (GRCz11:CM002902.2). The positions of the targeting Left and Right arms with a unique BamHI restriction endonuclease site in a spacer region between the TALEN pair are highlighted. b) Representation of the 11 bp deletion causing a premature stop codon. c) Graphical representation of the predicted Panx2 structure. The region in blue indicates the residual sequence of 27 amino acids followed by a new in-frame stop codon. Representative images (d) and comparison of body length between one-year-old zebrafish of different genotypes, both sexes. f) Real-time PCR quantification and presentation of the normalized expression of the three Panx2 isoforms, normalized to 18S rRNA. g) TL larvae showing that Panx2 proteins (green) are expressed throughout the retina. The expression was enriched in the outer retina, the inner plexiform layer, and the lens epithelium. DAPI is shown in magenta. h) After Panx2 ablation the expression was greatly reduced across the retina, with residual staining left in tissue known for high autofluorescence. i) Higher magnification of the outer retina showing the pronounced staining in sections of the inner photoreceptor segments, in TL larvae. j) Higher magnification of the lens epithelium of TL larvae. Abbreviations: SCL sclera, LE lens epithelium, GCL ganglion cell layer, IPL inner plexiform layer, INL inner nuclear layer, OPL outer plexiform layer, ONL outer nuclear layer, PRL photoreceptor layer. Scale bars: (g, h) 60 μm; (i, j) 10 μm. Statistics in e,f: Welch's test. Sample sizes were n = 26 for Panx2+/+ and n = 20 for Panx2Δ11. Significance: ***P-value<0.001 and *P-value<0.05. Error bars = SD. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)