Figure 6

Expression analysis of the candidate target genes in thalamic neurons (loss-of-function experiment). (A) Subcellular localization of β-catenin in thalamic neurons in vitro in control (Gfp-expressing; upper panel) and Axin2-expressing (lower panel) cultures. Neuronal marker NeuN is stained red. β-catenin is green, and nuclei are blue. The arrows point to nuclear β-catenin-positive neurons. Scale bar = 20 μm. The percentage of β-catenin-positive neurons in every culture is indicated, with p values of the differences (Fisher’s Exact test). (B) Expression of the candidate LEF1/TCF1 targets Cacna1g, Cacna2d2, Kcna6, Kcnh8, Drd3, Gabra3, Glra1, Grid2, and Calb2, neuronal marker Map2, negative control Cacna1h, and positive control Lef1 in thalamic cultures transduced with Axin2-expressing adenoviral vector compared with control (Gfp-expressing cultures). The expression levels are relative to the level of Gapdh. The graph shows the means of all of the results relative to the control, set at 1. Drd3 mRNA was not detected (ND). n = 9 independent biological samples. Error bars indicate SD. ***p < 0.001, **p < 0.01, *p < 0.05 (Student’s t-test).