Figure 4.

Immunostaining of the sagittal sections from the control and hausp^FL/FL; nes-cre brains at day E14.5. HAUSP was detected using a polyclonal antibody anti-HAUSP C terminus (a–d). HAUSP was ubiquitously expressed in all cells indicated by the presence of brown staining in nuclei of the control brain (a and b), whereas most of the cells from ventricular zone and intermediate zone of the hausp^FL/FL; nes-cre brain showed no staining of HAUSP protein (c and d). Immunostaining of p53 was done using an anti-p53 polyclonal antibody (CM5) (e–h). Dramatic p53 activation was detected in the hausp^FL/FL; nes-cre brain (g and h), whereas no p53 staining was detected in the control brain (e and f). Apoptosis was determined by the presence of Cleaved Caspase-3 (i–l). Consistent with the abundant appearance of pyknotic nuclei, Cleaved Caspase-3 was detected in large number of neural cells in the hausp^FL/FL; nes-cre brain by immunostaining (k and l). No Cleaved Caspase-3 was detected in the control brain (i and j). b, d, f, h, j, and l are the higher magnified pictures of the insects in a, c, e, g, i, and k, respectively. IZ, intermediate zone; MZ, marginal zone; V, ventricle; VZ, ventricular zone