Figure 7. Changes in L4 and L5 molecular markers in ThVGdKO mice.

(A) In situ hybridization for Rorb at P15 shows a dense band of labeled cells that is shifted toward the pial surface in ThVGdKO mice (right) relative to controls (left). Arrows mark sparse L5 cells, which are more prominent in ThVGdKO mice. Scale bar: 150µm. (B) Quantitative analysis of the laminar distribution of Rorb+ cells across layers (mean ± SEM) shows that the dense band of Rorb+ cells is shifted upward and there are an increased number of Rorb+ cells in deeper layers in ThVGdKO somatosensory cortex. The total number of Rorb+ cells is not different in the two genotypes (right panel). * P< 0.05; ***P< 0.001, n=4 for each genotype. (C) In situ hybridization for Etv1 at P15 in ThVGdKO (right) and littermate control (left) mice. Scale bar: 150µm. (D) The laminar distribution of Etv1+ cells in ThVGdKO mice is shifted toward the pial surface relative to control mice, and there are many more Etv1+ cells in ThVGdKO mice (mean ± SEM). *P< 0.05; **P< 0.01, n=5 for each genotype. (E) C-FOS expression is dramatically reduced in ThVGdKO mice (right) relative to control mice (left) at P14. (F) Laminar distribution of FOS expression (mean ± STDEV) shows reduced expression throughout cortex, particularly in superficial layers. ***P< 0.001; **P< 0.01, n=2. (G) EGR1 immunostaining in somatosensory cortex of ThVGdKO (right) and littermate controls (left) at P15. Scale bar: 300 µm. (H) EGR1 expression is reduced in ThVGdKO mice relative to littermate control mice, particularly in superficial layers (mean ± SEM). ** P< 0.01. (I) Western blot analysis shows that EGR1 and CUX1 expression are reduced in ThVGdKO somatosensory cortex, while ETV1 expression is increased. (J) Quantification of western blots for indicated proteins. * P< 0.05; ** P< 0.01, n=3 for controls and ThVGdKOs. See also Fig. S7.