FIGURE 4.

Neuronal morphology of cortical neurons overexpressing Cntn6, Lphn1 or in combination. (A) Mouse cortical cultures were transfected with Cntn6, Lphn1, Cntn6 + Lphn1 and control plasmids. Cultures were immunostained with an anti-GFP antibody (green). DAPI staining is in blue. (B) Morphological parameters of neurons cultured on Laminin were quantified and revealed a significant decrease on all parameters of the Lphn1 overexpressing neurons. (C) Representative photographs of caspase-3 (red) positive cells in neuronal cultures transfected with an EGFP expression vector (green) and vectors either for Cntn6, Lphn1, combination of both or an empty control plasmid. DAPI staining is in blue. Arrowheads indicate caspase-3 expression in the soma of neurons. (D) Apoptosis in neurons expressing Cntn6 and/or Lphn1 was quantified, showing an increase of apoptosis in Lphn1-expressing neurons. This was reduced by coexpression with Cntn6. About 60 transfected neurons were analyzed for caspase-3 immunoreactivity per condition of each independent experiment (n = 3). The scale bars indicate 100 μm. Statistical analyses were performed using unpaired Student’s t. The graph bars are presented as mean ± SEM. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001.