Nbn and Atm Cooperate in a Tissue and Developmental Stage-Specific Manner to Prevent Double Strand Breaks and Apoptosis in Developing Brain and Eye
Figure 5
Inactivation of Nbn and Atm leads to precocious DSBs accumulation and apoptosis in embryonic lens.
(A) BrdU immunohistochemistry of NbnCtrl, NbnNes-Cre and Nbn/AtmNes-Cre lens at E15.5 (Magnification ×200). (B) BrdU immunohistochemistry of NbnCtrl, NbnNes-Cre and Nbn/AtmNes-Cre lens at E17.5 (Magnification ×200). (C) Quantification of BrdU positive cells within lens anterior epithelia at E15.5 (NbnCtrl, n = 3; NbnNes-Cre, n = 3; and Nbn/AtmNes-Cre, n = 3) and E17.5 (NbnCtrl, n = 7; NbnNes-Cre, n = 6; and Nbn/AtmNes-Cre, n = 3). (D) γ-H2AX immunohistochemistry of NbnCtrl, NbnNes-Cre and Nbn/AtmNes-Cre lens at E15.5 (Magnification ×1000). (E) γ-H2AX immunohistochemistry of NbnCtrl, NbnNes-Cre and Nbn/AtmNes-Cre lens at E17.5 (Magnification ×1000). (F) Quantification of γ-H2AX positive cells within lens anterior epithelia at E15.5 (NbnCtrl, n = 3; NbnNes-Cre, n = 3 and Nbn/AtmNes-Cre, n = 5) and E17.5 (NbnCtrl, n = 3; NbnNes-Cre, n = 3 and Nbn/AtmNes-Cre, n = 3). (G) TUNEL staining of NbnCtrl, NbnNes-Cre and Nbn/AtmNes-Cre lens at E15.5 (Magnification ×400). (H) TUNEL immunohistochemistry of NbnCtrl, NbnNes-Cre and Nbn/AtmNes-Cre lens at E17.5 (Magnification ×400). (I) Quantification of TUNEL cells within lens anterior epithelia at E15.5 (NbnCtrl, n = 20; NbnNes-Cre, n = 2 and Nbn/AtmNes-Cre, n = 5) and E17.5 (NbnCtrl, n = 7; NbnNes-Cre, n = 3 and Nbn/AtmNes-Cre, n = 5). DSB accumulation and apoptotic cell death occurs earlier in Nbn/Atm-deficient lens as compared to Nbn-deficient lens. (J) Phospho-histone 3 (H3-P) immunohistochemistry of NbnCtrl, NbnNes-Cre and Nbn/AtmNes-Cre lens at E15.5 (Magnification ×400). (K) Phospho-histone 3 (H3-P) immunohistochemistry of NbnCtrl, NbnNes-Cre and Nbn/AtmNes-Cre lens at E17.5 (Magnification ×400). (L) Quantification of Phospho-histone 3 positive cells (H3-P+) cells within lens anterior epithelia at E15.5 (NbnCtrl, n = 6; NbnNes-Cre, n = 5 and Nbn/AtmNes-Cre, n = 2) and E17.5 (NbnCtrl, n = 13; NbnNes-Cre, n = 3 and Nbn/AtmNes-Cre, n = 5). Error bars indicate SEM. (* p<0,05, ** p<0,01, *** p<0,001, ns: non-significant).