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

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© 2013 Rodrigues et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Body weight of P9 Nbn/Atm^F/F;Nestin-cre (Nbn/Atm^Nes-Cre) mice was reduced as compared to control (Nbn^Ctrl) and Nbn^F/F;Nestin-Cre (Nbn^Nes-Cre) littermates. (B–C) Brain weight (B) and size (C) of P9 of Nbn^Nes-Cre (n = 11) and Nbn/Atm^Nes-Cre (n = 22) mice are significantly reduced as compared to those of Nbn^Ctrl (n = 123), without dramatic disruption of the general morphology. (D) H&E staining of Nbn^Ctrl, Nbn^Nes-Cre and Nbn/Atm^Nes-Cre brain sections at P9 reveal reduction of the size of the Nbn^Nes-Cre and Nbn/Atm^Nes-Cre mice cerebella compared to those of Nbn^Ctrl mice (Magnification ×10). (E) Calbindin (D-28K) staining shows disruption of the Purkinje cells layer in both Nbn^Nes-Cre and Nbn/Atm^Nes-Cre cerebella (Magnification ×400). (F) NeuN staining shows the disorganization of the inner granular layer (IGL) and the dramatic size reduction of the external granular layer (EGL) in Nbn^Nes-Cre and Nbn/Atm^Nes-Cre cerebella (Magnification ×400). (G) Purkinje cells number is unchanged in both Nbn^Nes-Cre (n = 5) and Nbn/Atm^Nes-Cre (n = 3) cerebella compared to Nbn^Ctrl (n = 5) cerebella. (H) Inactivation of Nbn (Nbn^PcP2-Cre; n = 2) or both Nbn and Atm (Nbn/Atm^PcP2-Cre; n = 2) in post-mitotic Purkinje cells using PcP2-Cre transgene does not alter the number of Purkinje cells in adult cerebellum. (I) Genetic inactivation of Nbn and Atm in the lens epithelia severely impairs eye and lens development. Representative pictures of P9 eyes from Nbn^Ctrl, Nbn^Nes-Cre and Nbn/Atm^Nes-Cre mice. (J) Representative pictures of P9 dissected lenses from Nbn^Ctrl, Nbn^Nes-Cre and Nbn/Atm^Nes-Cre. (K) Reduction in volume of the Nbn-deficient eyes was potentiated by simultaneous inactivation of both Nbn and Atm. Nbn^Ctrl (n = 17), Nbn^Nes-Cre (n = 14) and Nbn/Atm^Nes-Cre (n = 28). (L) Lens volume measurements show that simultaneous inactivation of Nbn and Atm results in a greater reduction of lens volume as compared to control or Nbn-deficient lens, Nbn^Ctrl(n = 7), Nbn^Nes-Cre (n = 6) and Nbn/Atm^Nes-Cre (n = 15). Error bars indicate SEM. (* p<0,05, ** p<0,01, *** p<0,001, ns: non-significant).