<p><b>A-P</b>: Magnifications from SD flat mounted retinas analyzed 3 and 5 days after intraorbital nerve crush (<b>A–H</b>) and intraorbital nerve transection (<b>J–P</b>). As internal control, Brn3a was double immunodetected with Brn3b (<b>A–B, E–F, I–J, M–N)</b> or with Brn3c (<b>C–D, G–H, K–L, O–P</b>). In these images is observed that Brn3b and Brn3c expression decreases already 3 days after both injuries. This low signal impeded the automated quantification of Brn3b and Brn3c positive RGCs. Brn3a<sup>+</sup>RGCs were counted in these retinas and isodensity maps showing the distribution of the surviving RGCs were created (<b>Q–T</b>, at the bottom of each map is shown the number of Brn3a<sup>+</sup>RGCs counted in the retina wherefrom the map has been generated). These maps illustrate that IONT induces a quicker loss of RGCs than IONC, as evidenced by the higher densities observed in <b>Q</b> and <b>S</b> than in <b>R</b> and <b>T</b>. Colour scale (T bottom right) is the same as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0049830#pone-0049830-g004" target="_blank">figure 4M</a>. Four (IONC 5d) or five (rest of the groups) retinas were analyzed per marker. <i>Bars:</i> 50 µm (J), 1 mm (Q).</p

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