BUSCADOR RECOLECTA

Experimental Procedures for Supplemental Figure: Double-stranded Zic2 and Zic2 mutant decoy DNA were prepared by annealing complementary single strands with the follow sequences: 5′-TCTTGGGTGGTCTCCGGAGACCACCCAAGA (for Zic2 decoy) and 5′-TCTTGAGTGAACTCCGGAGTTCACTCAAGA (for Zic2M decoy) as a negative control (Mizugishi et al., 2001). We utilized unlabeled and also Texas-red-tagged oligonucleotides (Invitrogen) to monitor DNA entry into cells. The terminal four bases on either site of the molecule were linked by phosphorothioate esters for added stability. Retinal explants were exposed to serial concentrations (0, 10, 100, and 200 μM) of decoy DNA combined with serial concentrations of serum-free medium (SFM) plus Fungene 6 transfection reagent (Roche) and incubated for 1 hr 30 min at 37°C. The explants were then washed several times in SFM, plated on D-polylysine/laminin, and incubated at 37°C, and dissociated chiasm cells were added to the cultures 2 hr later. They were then immunolabeled with α-neurofilament and explant outgrowth quantified, as indicated in Experimental Procedures., Figure S1. Zic2 Is Necessary for Ventrotemproal (VT) RGCs to Respond Negatively to Cues from the Chiasm Midline In Vitro. (A) Retinal explants grown on dissociated chiasm cells and immunostained with α-neurofilament. Representative images of dorsotemporal (DT) (a–d) and VT (e–h) retinal explants that were untransfected (a and e) or incubated with lipofectamine (b and f), Zic2 decoy oligos (c and g), or Zic2-mutant-decoy oligos (d and h) and subsequently cocultured with dissociated chiasm cells. Scale bar: 100 μm. (B) Quantification of area occupied by axons of DT (black columns) or VT (white columns) retinal explants that were untransfected or incubated with lipofectamine, Zic2-decoy oligos, or mutant-Zic2-decoy oligos, and then cocultured with dissociated chiasm cells. Left panels and graph: transfection of Zic2-decoy oligos does not affect the outgrowth in DT neurites (p > 0.2). Right panels and graph: transfection of Zic2-decoy oligos leads to increased growth of VT neurites. (*p < 0.05 compared with VT explants incubated with the lipofection agent alone or transfected with Zic2M-decoy). Number above bars indicates number of explants., Peer reviewed

Analytical and spectral characterization data of all new compounds, NMR/NOESY interprotonic distances, and crystallographic data (CIF)., Peer reviewed

La Rábida. Revista Colombina Iberoamericana, año VII, nº 72, 30 de junio de 1917, pág. 13. Fotografía blanco/negro., Pie de foto: Parroquia de San Juan Bautista, La Palma, Huelva, http://hdl.handle.net/10334/1413

La Rábida. Revista Colombina Iberoamericana, año VII, nº 74, 31 de agosto de 1917, pág. 11. Fotografía blanco/negro., Pie de foto: Monasterio de La Rábida, Huelva, los muebles donados por los Marqueses de Aracena, colocados en las celdas de la Colombina, http://hdl.handle.net/10334/1415

La Rábida. Revista Colombina Iberoamericana, año VII, nº 69, 31 de marzo de 1917, pág. 5. Fotografía blanco/negro., Pie de foto: Otro detalle de las ruinas del castillo romano, Ayamonte, Huelva, http://hdl.handle.net/10334/1410

La Rábida. Revista Colombina Iberoamericana, año VII, nº 73, 31 de julio de 1917, pág. 5. Fotografía blanco/negro., Pie de foto: Plaza de la Constitución, Cartaya, Huelva, http://hdl.handle.net/10334/1414

La Rábida. Revista Colombina Iberoamericana, año VII, nº 74, 31 de agosto de 1917, pág. 8. Fotografía blanco/negro., Pie de foto: Pedro Garrido Perelló, Presidente de la Comisión de Fiestas, http://hdl.handle.net/10334/1415

La Rábida. Revista Colombina Iberoamericana, año VII, nº 71, 31 de mayo de 1917, pág. 5. Fotografía blanco/negro., Pie de foto: Vista de la Plaza de la Constitución viéndose al fondo el Ayuntamiento, Cartaya, Huelva, http://hdl.handle.net/10334/1412

La Rábida. Revista Colombina Iberoamericana, año VII, nº 74, 1 de agosto de 1917, pág. 10. Fotografía blanco/negro., Pie de foto: Monasterio de La Rábida, Huelva, puerta exterior de la iglesia cuya restauración ha comenzado en estos días, http://hdl.handle.net/10334/1415

La Rábida. Revista Colombina Iberoamericana, año VII, nº 69, 31 de marzo de 1917, pág. 13. Fotografía blanco/negro., Pie de foto: D. Benito Pérez Galdós (X) rodeado de los catedráticos y alumnos del Instituto, http://hdl.handle.net/10334/1410