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. 2020 Mar 16;130(4):2054–2068. doi: 10.1172/JCI125898

Figure 1. cAMP induces neogenin expression in the retina.

Figure 1

(A and B) Retinoblastoma (Y79) cells were cultured with or without 0.5 mM 8-Br-cAMP (cAMP) or 0.5 mM 8-Br-cGMP (cGMP) for 48 hours. (A) Western blot of neogenin from Y79 whole cell lysates following the indicated treatment. Ctrl, Control. (B) Quantification of data from A, normalized to GAPDH (n = 3 for each; *P < 0.05, **P < 0.01). Significance determined by 1-way ANOVA followed by Šidák’s multiple-comparisons test. (C–E) P9 C57BL/6J (WT) mice received 1 intravitreal injection of either 10 mM 8Br-cAMP or PBS. Eyes were harvested on P12 and cryosectioned. (C) Representative images of P12 retina treated with PBS or 10 mM 8Br-cAMP and stained for neogenin (red), rhodopsin (green), and nuclei (DAPI; blue). Arrows indicate inner/outer segments of photoreceptors. IPL, inner plexiform layer. Scale bars: 20 μm. (D) Higher-magnification images of P12 retina treated with PBS or 8Br-cAMP stained for neogenin (red), with the dashed lines indicating the border of the inner/outer segments (IS/OS). Note the characteristic punctate appearance of neogenin staining. Scale bar: 20 μm. (E) Quantification of data from D. Neogenin signal in the IS/OS was significantly elevated following 8Br-cAMP intravitreal injection (n = 4 for each; *P < 0.05). Significance determined by Student’s t test.