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. 2013 Mar 8;6(3):768–779. doi: 10.1242/dmm.010397

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© 2013. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial Share Alike License (http://creativecommons.org/licenses/by-nc-sa/3.0), which permits unrestricted non-commercial use, distribution and reproduction in any medium provided that the original work is properly cited and all further distributions of the work or adaptation are subject to the same Creative Commons License terms.

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Fig. 4. — Maxillary-palatine suture at P0. (A,B) H&E staining shows a similar overlap of maxillary (max) and palatal (pal) bones in unaffected littermates (Fgfr2^+/+) and Fgfr2^+/S252W sutures, but secreted osteoid bridges the two bones (arrowhead) in a mutant suture beginning to fuse. (C,D) Osteoblast-specific ALP activity is absent from normal suture mesenchyme (C, arrow). In the early stages of fusion, ALP is still absent in mutant mesenchyme (D, arrow) dorsal to the point of fusion (white arrowhead), but has risen throughout mesenchyme ventral (black arrowhead) to this point (D). ALP activity is imaged fluorescently and shown in gray channel. (E,F) Immunofluorescent staining for Ki67 (green) shows a similar frequency of positive cells in the suture mesenchyme and adjacent osteogenic surfaces (stained for ALP activity; red) of the maxillary and palatal bones between unaffected littermates and mutant mice. (G,H) Immunofluorescent staining for Runx2 (green) shows similar expression levels between unaffected littermates and mutant mice, with expression being generally lower in the suture mesenchyme than in osteoblasts within osteogenic fronts (stained for ALP activity; red). (I,J) TUNEL staining (green) shows that apoptosis was negligible within the suture mesenchyme and infrequent in the bones of wild-type sutures. In mutant sutures, apoptosis was intense specifically in areas undergoing fusion (arrowhead). (K,L) Phospho-p38 levels within bone and suture mesenchyme were similar in mutant and unaffected littermate sutures, although expression was decreased in the area of mutant suture dominated by apoptotic cells (arrowhead). (M,N) Phospho-ERK 1/2 expression was concentrated around bone at similar levels in mutant and unaffected littermate sutures. (O) The percentage of suture mesenchyme cells expressing Ki67 was not significantly different between mutant and unaffected littermates. All images are from near-adjacent parasagittal sections from the same littermate pair. Rostral direction is to the right. Scale bar: 100 μm.