a, b) GFAP immunostaining (green) in coronal sections containing electroporated neurons (red) under control ( a) or Rheb CA condition ( b). The dotted line and white arrows point to a bulging of the ACC containing Rheb CA-expressing cytomegalic neurons. Scale bar, 150 μm. ( c) Bar graphs of fluorescence (Fu) intensity of GFAP immunoreactivity in both the ipsilateral (ipsi) and contralateral (contra) cortex under conditions shown in ( a, b). P<0.01 (two-way analysis of variance (ANOVA)); * P<0.05, ** P<0.01 (Sidak's multiple comparisons post-test). ( d) T1-weighted MR images of control ( d) and Rheb CA ( e) brains showing measuring bars for cortical thickness. ( e) Quantification of cortical thickness shown in d, e with N=6 per condition. P<0.001 (two-way ANOVA); *** P<0.001 (Sidak’s multiple comparisons post-test). ( f) Serial tomograms from diffusion tensor imaging (DTI) with statistically significant ( P<0.01) differences in fractional anisotropy (FA; Rheb CA minus control, expressed in heat maps with scale at the bottom) between control ( N=4) and Rheb CA ( N=4) transfected brains, overlaid on top of anatomical scans of a control brain. ( g) Top, GABA immunostaining in coronal sections from mice electroporated with control or Rheb CA plasmid. Scale bar, 300 μm. Bottom, bar graph of the density of GABAergic cells in control and Rheb CA-expressing sections. Error bars, s.e.m.