Open Access Highly Accessed Research article

Reprogramming of the chick retinal pigmented epithelium after retinal injury

Agustin Luz-Madrigal12, Erika Grajales-Esquivel1, Alexander McCorkle1, Ashley M DiLorenzo1, Karla Barbosa-Sabanero1, Panagiotis A Tsonis2 and Katia Del Rio-Tsonis1*

Author Affiliations

1 Department of Biology, Miami University and Center for Visual Sciences at Miami University (CVSMU), Oxford, OH 45056, USA

2 Department of Biology, University of Dayton and Center for Tissue Regeneration and Engineering at the University of Dayton (TREND), Dayton, OH 45469, USA

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BMC Biology 2014, 12:28  doi:10.1186/1741-7007-12-28

Published: 17 April 2014

Additional files

Additional file 1: Figure S1:

Pluripotency inducing factors are present in the developing chick eye at E7 and after injury. (A-E) Immunohistochemical staining using antibodies against pluripotency inducing factors c-Myc (A), Sox2 (B), Klf4 (C), Lin‒28 (D) and progenitor markers Pax6 and Chx10 (E) in the posterior region of E7 (this stage was included for comparison with the 72 h PR, as the developing retina would be equivalent to the regenerating retina 72 h PR) chick eyes. Higher magnification view of the boxed areas are shown in A’-E’; dashed lines outline the RPE. (F-J) Immunofluorescence analysis of c‒Myc (F), Sox2 (G), Klf4 (H), Lin-28 (I) and progenitor markers Pax6 and Chx10 (J) in eyes 72 h PR. (K-M) Negative controls (only secondary antibodies added) for immunofluorescence analysis on eyes 72 h PR for Sox2 and Lin-28 (K), c‒Myc and Klf4 (L) and Pax6 and Chx10 (M). L, lens; M, mesenchyme; R, retina; RPE, retinal pigmented epithelium; Asterisk, FGF2-soaked heparin bead. The scale bar in panel E represents 50 μm and applies to panels A-E. Scale bar in panel E’ represents 10 μm and applies to panels A’-E’. The scale bar in panel J represents 50 μm and applies to panels F-J. The scale bar in panel M represents 50 μm and applies to panels K-M.

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Additional file 2: Figure S2:

FGF2 induces proliferation of the RPE. (A-H) Retinectomies were performed on E4 (Stage 24) chick eyes in the absence (A-D) or presence (E-H) of FGF2. Immunostaining using an antibody to detect BrdU (A,C,E,G), shows BrdU + RPE cells only at 24 h post-retinectomy (PR) in the presence of FGF2. Immunostaining to detect p27kip1 (B,D,F,H) is negative in the RPE only at 24 h PR in the presence of FGF2. L, lens; M, mesenchyme; RPE, retinal pigmented epithelium; *Asterisk, FGF soaked heparin bead. The scale bar in panel H represents 50 μm and applies to all images.

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Additional file 3: Table S1:

Primer sequences for RT-PCR. Table S2. Primer sequences for RT-qPCR. Table S3. Primer sequences for lin-28 cloning. NCBI accession number NM_001031774.

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