Fig. 2

Upper layer defects in Zbtb20 ^lacZ/lacZ mice. a1-b2 Cresyl violet staining of brain cross sections from P28 wild type (WT) and mutant cortex. (b1, b2) are images at high magnification from fields in the somatosensory cortex, labeled with * in A1/A2. (b3-b4) NeuN IHC at stage P4. c1-g4 Evaluation of UL neuronal fate by using ROR and Cux1 antibodies as markers, and a statistical analysis. In (c1-c2) note the expansion of the ROR signal (arrows), while in (d1-d2), a slight reduction of the Cux1-positive band (arrows) in the mutant Ncx. (e1-g2) are higher magnifications of SS cortex, labeled with the same markers. (c3,d3,g3,g4) represent graphs of statistical analysis of the number of positive cells/frame in SS cortex (*, P < 0.05, n = 3 per genotype). h1-i2 IHC with Brn2 and Satb2 antibodies as UL markers on cross sections from P4 brains, and a statistical analysis of the number of stained cells. Note the reduction of the ULs in the mutant, which was statistically confirmed (*, P < 0.05, n = 3 per genotype). j1-j3 Double IHC with Brn2 and FoxP1 antibody to evaluate UL neurons with L2 identity (Brn2⁺/FoxP1⁻). Note the drastic diminishing of the number of L2 neurons in the mutant Ncx (arrows), which is supported by a statistical analysis (*, P < 0.05, n = 3 per genotype). k1-k2 Double immunostaining for ROR (L4) and Ctip2 (L5) reveals a preserved molecular border between the two layers in the mutant. Countings of positive cells were performed in frames sized 300 μm (h) × 100 μm (w) spanning the ULs of SS cortex. Scale bars: b4, 100 μm; d2, 500 μm; g2/j2, 50 μm; i2/k2, 100 μm