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. 2013 Jul 30;8(7):e69209. doi: 10.1371/journal.pone.0069209

Figure 3. Nbn and Atm inactivation leads to accumulation of DSBs, proliferation defects and apoptosis in the EGL of the cerebellum.

Figure 3

(A) Loss of Nbn and Atm leads to increase of DSBs at E15.5 as indicated by γ-H2AX foci (NbnCtrl, n = 3; NbnNes-Cre, n = 5 and Nbn/AtmNes-Cre, n = 4) (Magnification ×400). (B) Accumulation of neural progenitors in mitosis in EGL primordium as indicated by H3 phosphorylation (Magnification ×400). (C) Cleaved Caspase-3 staining demonstrates increased apoptosis in NbnNes-Cre and Nbn/AtmNes-Cre EGL. (Magnification ×400) (D) BrdU immunohistochemistry shows the reduction in cell proliferation of the neural progenitors in the NbnNes-Cre and Nbn/AtmNes-Cre EGL (Magnification ×400). (E) TUNEL staining demonstrates increased apoptosis in NbnNes-Cre and Nbn/AtmNes-Cre EGL (Magnification ×400). (F) p53 is stabilized in NbnNes-Cre and Nbn/AtmNes-Cre EGL (Magnification ×400). (G) Quantification of cells with γ-H2AX foci in the EGL primordium at E15.5 (NbnCtrl, n = 3; NbnNes-Cre, n = 5 and Nbn/AtmNes-Cre, n = 4) and E17.5 (NbnCtrl, n = 3; NbnNes-Cre, n = 3 and Nbn/AtmNes-Cre, n = 3). (H) Quantification of phospho-histone 3 positive cells (H3-P+) in the EGL primordium at E15.5 (NbnCtrl, n = 4; NbnNes-Cr e, n = 5 and Nbn/AtmNes-Cre, n = 5). (I) Quantification of BrdU positive cells in the EGL primordium At E15.5 (NbnCtrl, n = 5; NbnNes-Cre, n = 3 and Nbn/AtmNes-Cre, n = 4). (J) Quantification of TUNEL positive cells in the EGL primordium at E15.5 (NbnCtrl, n = 4; NbnNes-Cre, n = 3 and Nbn/AtmNes-Cre, n = 4) and E17.5 ((NbnCtrl, n = 3; NbnNes-Cre, n = 3 and Nbn/AtmNes-Cre, n = 4). (K) Quantification of Cleaved Caspase-3 positive cells in the EGL primordium at E15.5 (NbnCtrl, n = 3; NbnNes-Cre, n = 5 and Nbn/AtmNes-Cre, n = 4) and E17.5 (NbnCtrl, n = 3; NbnNes-Cre, n = 3 and Nbn/AtmNes-Cre, n = 4) Error bars indicate SEM. (* p<0,05, ** p<0,01, *** p<0,001, ns: non-significant).