Fig. 1

Conditional overexpression of KCC2 in motor cortex in transgenic mice. a Representative Western blots for KCC2 immunoreactivity in tissue from the motor cortex obtained from three WT (left) and KCC2-overexpressing (right) mice. Samples were obtained from mice 7 days after withdrawal of doxycycline (Dox) from the diet to induce overexpression in KCC2 mice. b Quantification of the intensity of KCC2 immunoreactivity, expressed relative to β-actin. The relative expression of KCC2 was significantly greater in KCC2-overexpressing mice compared with WT mice, with the absolute KCC2/β-actin ratios being 40.4 ± 9.3% for WT (n = 3) and 87.8 ± 9.8% for KCC2 mice (n = 3; unpaired t test,*p < 0.05). This corresponded to a 117% increase in KCC2 protein expression levels. c Representative immunohistochemical images of a brain slice from a KCC2-overexpressing transgenic mouse showing the primary motor cortex (left) stained positive for NeuN (red, neuronal marker) and VP16 (green, marker for the tetO-KCC2 construct). Most larger layer V pyramidal neurons were double-labeled (yellow). Bar; 500 μm (left), 10 μm (upper right) and 50 μm (bottom right)