RA and FA co-supplementation rescue Wnt signaling.

<p>Alcama staining on <i>Tg(Tcf/lef-miniP</i>:<i>dGFP)</i> fish showed that ethanol treatment reduced the number of Wnt+/Alcama+ cells in the peripheral CMZ. Control and ethanol treated embryos in this experiment appeared identical to those shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0201659#pone.0201659.g004" target="_blank">Fig 4A–4C</a> and were left out for brevity. (A-F) RA co-supplementation could rescue Wnt+/Alcama+ cells (A-C); FA co-supplementation could also rescue Wnt+/Alcama+ cells in the CMZ particularly in 100 mM Ethanol + FA treated fish (D-F). (G) Quantification of Wnt+/Alcama+ cells in the peripheral CMZ showed significant reduction after ethanol treatment, which was rescued by RA and FA co-supplementation. Numbers of cells were counted in a single optic-nerve containing confocal optical section in the ventral CMZ. Images show rostral at top, lateral at left. Error bars indicate standard deviation. ‘**’ indicates statistical significance in comparison to control embryos (p<0.01). ‘*’ indicates statistical significance in comparison to control embryos (p<0.05). ‘a’ indicates statistical significance in comparison to 100 mM ethanol embryos (p<0.05). ‘b’ indicates statistical significance in comparison to 150 mM ethanol embryos (p<0.05). Scale bar = 10 μm for panels A-F.