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. 2007 Jul 9;204(7):1613–1623. doi: 10.1084/jem.20062525

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Copyright © 2007, The Rockefeller University Press

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Figure 5. — ATM functions in the development of growth plates. (A) Tibial sections of 3-wk-old wild-type mice were stained with rabbit antiphosphorylated ATM (pATM) antibody, followed by Alexa Fluor 488–conjugated anti–rabbit Ig antibody, and observed under a confocal microscope. Nuclei were stained with TOTO3. Representative data are shown. Growth plates are indicated between the dotted lines. pATM was detected in the chondrocytes of growth plates. Bar, 100 μm. (B) Increased intracellular ROS was detected by dihydroethidium staining in growth-plate chondrocytes of 2-wk-old Atm^−/− mice compared with those seen in wild-type (Atm^+/+) mice. Bar, 100 μm. (C) Chondrocyte proliferation was evaluated by BrdU staining in 2-wk-old Atm^−/− or wild-type (Atm^+/+) mice. (left) Representative data are shown. (right) Data are mean relative numbers ± SD of BrdU-positive cells in Atm ^−/− compared with Atm^+/+ chondrocytes. Growth plates are shown between the dotted lines. Atm ^−/− deficiency caused defective chondrocyte proliferation. *, P < 0.01. Bar, 50 μm. (D) Disruption of columnar formation in proliferating zones (P) and enlargement of hypertrophic zones (H) were observed in growth plates of 3-wk-old Atm ^−/− compared with Atm^+/+ mice by H&E (top) and Alcian blue (bottom) staining. Bar, 50 μm. (E) Mineralization was evaluated by alizarin red staining in growth plates of 3-wk-old Atm ^−/− or wild-type (Atm^+/+) mice. Frozen sections that had not been decalcified were stained with alizarin red. Bar, 100 μm.