A. EdU staining for DMSO and 1 μM PR1 treated retinas explanted at P0. ... A. Staining for GFAP on sections from wild type retina explanted at P12 and treated ...
Supplemental Figure 1 Structures of Photoregulin1 (PR1) analogs PR101 to PR116. Supplemental Figure 2 Typhoon scan of 1 μM PR101-116-treated dissociated retinal cultures stained for Rhodopsin and Otx2. Supplemental Figure 3 A. EdU staining for DMSO and 1 μM PR1 treated retinas explanted at P0. Scale bar represents 50 μm. B. Schematic and cell counts for P0 retinal explants treated for 3 days in vitro (DIV) with DMSO or 1 μM PR1. EdU was included in the media for the last day. PR1 treatment did not affect the number of EdU-positive cells counted over 425 μm of central retina (n = 6, p > 0.05, Student’s t-test).
Supplemental Figure 4 A. Staining for GFAP on sections from wild type retina explanted at P12 and treated with media containing DMSO or 1 μM PR1 for 5 days in vitro (DIV). B. P7 wild type retinas were cultured for 2 DIV with DMSO or 1 μM PR1 and analyzed by Western blot for GFAP. C. GFAP expression was normalized to β-actin expression. PR1 did not cause an increase in GFAP (n = 4, p > 0.05, Student’s t-test). D. Staining for GFAP in sections from Pde6brd1 retinas explanted at P7 in media containing DMSO or 1 μM PR1 for 14 DIV. E. P7 Pde6brd1 retinas treated for 12 DIV with DMSO or 1 μM PR1 were analyzed for GFAP with Western blot analysis. F. GFAP expression was normalized to β-actin expression. PR1 did not cause an increase in GFAP (n = 3, p > 0.05, Student’s t-test). Scale bar in Panel D represents 50 μm and also applies to Panel A.
Supplemental Figure 5 A. Western blot for TRβ2 shows that intravitreal injection of PR1 increases expression compared to the uninjected, contralateral retina of adult mice. B. TRβ2 expression was normalized to β-actin expression. PR1 increased the relative expression of TRβ2 after intravitreal injection in adult mice (n = 5, *p < 0.05, Student’s t-test).
Supplemental Figure 6 A. Staining for Rhodopsin and S Opsin on sections from adult (P60) retinas explanted in media with DMSO or 1 μM PR1 for 2 days in vitro (DIV). PR1 caused a decrease in Rhodopsin expression. B. GFAP and Sox2 staining on sections from adult (P60) retinas treated with DMSO or 1 μM PR1 for 2 days. PR1 treatment did not result in increased GFAP staining nor migration of Sox2-positive nuclei to the outer nuclear layer (ONL). Scale bar in Panel B represents 50 μm and also applies to Panel A.
Supplemental Figure 7 A. TUNEL staining (green, arrows) on retinal sections from P7 Pde6brd1 treated for 9 days in vitro with DMSO or 1 μM PR1. Scale bar represents 50 μm. B. PR1 treatment decreased the number of TUNEL-positive nuclei in the outer nuclear layer (ONL) of Pde6brd1 retinas explanted at P7 (n = 4, *p < 0.05, Student’s t-test).
