Aberrant cortex depth and ventricular zone cells in Nfix -/- mice. Brains from WT (+/+) and Nfix-/- animals were harvested, fixed in 4% PFA and imaged before (A) or after (B-E) sectioning, staining and mounting. A) Gross structure of P22 brains of WT (+/+) and Nfix -/- animals. Note difference in cortex anterior-posterior length (double-headed arrows). B) Cresyl violet-stained coronal sections of P22 WT (+/+) and Nfix -/- brains. Arrows denote normal ventricular region in Nfix+/+ and aberrant ventricular zone cells in Nfix-/- brains. C) Immunostaining of Pax6 in coronal sections of P16 WT (+/+) and Nfix -/- brains. Arrow shows Pax6 expression in aberrant ventricular zone cells of an Nfix-/- brain and in the ventricular zone of a +/+ littermate brain. D) Pax6 and phospho-histone H3 staining of the ventricular region. Pax6 (α-Pax6, panels 1&3) and phospho-histone H3 (α-pH3, panels 2&4) immuno-staining of ventricular regions of P16 WT (+/+, panels 1&2) and Nfix-/- (-/- panels 3&4) brains. Arrows in panels 2&4 denote some pH3-positive cells. Note that most of the Pax6 positive ventricular cells in the -/- brains do not react with α-pH3. Bar in panel 1 = 100 μm for D1-4 and 500 μm for both panels of C. E) Doublecortin (DCX) immuno-staining in p69 WT (+/+) and Nfix -/- brains. Panels 2&4 are higher magnification images of the boxed regions of panels 1&2, respectively. Arrows show the locations of some DCX-positive cells. Bars in panels 2&4 = 50 μm and in panel 3 = 2 mm.