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

The wide-angle seismic reflection data set consists of two transects. Transect A coincides with the trace of the IBERSEIS deep seismic reflection profile and transect B is located farther to the SE. The explosive charge was located in a single 20 cm diameter, 50–60-m deep shothole. Explosive shots with charge sizes of 1000, 750 and 500 kg, where distributed along the transect, with the largest charges located at the edges of the profile. 650 digital seismic recorders (590 Texans and 60 3-component Refteks) from IRIS (Incorporated Research Institutions for Seismology) instrument pool where used. These instruments where placed along the 300-km transect A at a 400-m station spacing. For transect B, a denser trace spacing was used. To achieve this, shots were fired twice. The stations were firstly placed at 300–400 m spacing along transect B. Once the shots were fired, the stations were moved 150–200 m toward the north along the profile. All shots were then fired a second time, resulting in shot records with a 150–200 m trace spacing., The IBERSEIS wide-angle seismic reflection dataset consists of two NE-SW oriented transects acquired in the SW of the Iberian Massif. They crossed three major geological zones (South Portuguese Zone, Ossa-Morena Zone, and Central Iberian Zone), with their tectonic contacts and the Pyrite Belt being of greatest interest. The velocity models obtained by forward modeling show a complex crust, especially in the middle crust. The velocity models are the most detailed ones that have been produced in the area and contain a large amount of new features that are relevant to the understanding of the composition of the crust and upper mantle beneath the zone., Spanish Ministry of Science and Innovation, CGL2004-04623, CSD2006-00041.-- Generalitat de Catalunya.-- Junta de Andalucía.--, Peer reviewed