1999; 14:1734741. Zhang J, Tan X, Li W, Wang Y, Wang J, Cheng
1999; 14:1734741. Zhang J, Tan X, Li W, Wang Y, Wang J, Cheng X, Yang X. Smad4 is required for the normal organization of the cartilage growth plate. Dev Biol. 2005; 284:31122. [PubMed: 16023633]Author Manuscript Author Manuscript Author Manuscript Author ManuscriptDev Biol. Author manuscript; available in PMC 2016 April 01.Lim et al.PageHighlights Deletion of Smad4 or form I Bmp receptors in limb bud mesenchyme abolishes appendicular skeleton Loss of Smad4 prevents precartilaginous mesenchymal condensation within a cellautonomous manner Smad4 deletion does not impair expression of Ncad, Ncam1 and Ncam2 in limb mesenchymal cells Forced-expression of Sox9 doesn’t restore limb skeleton inside the absence of SmadAuthor Manuscript Author Manuscript Author Manuscript Author ManuscriptDev Biol. Author manuscript; out there in PMC 2016 April 01.Lim et al.PageAuthor Manuscript Author Manuscript Author Manuscript Author ManuscriptDev Biol. Author manuscript; accessible in PMC 2016 April 01.Figure 1. Conditional deletion of Smad4 or Alk2/3/6 in the limb mesenchyme causes CDK5 Inhibitor Synonyms severe skeletal defects(A) Gross morphology of wild form (WT) or Prx1-Cre; Smad4f/f (PS4) mice at birth. (B-F) Whole-mount skeletal staining of newborn mice using the genotype of wild form (B), Prx1Cre; Smad4f/f (C), Prx1-Cre; Alk3f/- (D), Prx1-Cre; Alk3f/-; Alk6+/- (E) or Prx1-Cre; Alk2f/-; Alk3f/-; Alk6+/- (F). (B’-F’) Greater magnification on the sternum region in the corresponding skeleton above. Arrows denote defects at the skull, sternum and hindlimb. (G) Whole-mount skeletal staining of E16.5 embryos together with the genotype of wild kind (WT) or Prx1-Cre;Alk3f/- (PA3). (H) H E staining of a longitudinal section via the humerus of the wild-type (WT) or the forelimb of the Prx1-Cre; Alk2f/-; Alk3f/- littermate embryo (PA23) at E16.five. (I) H E staining of a longitudinal section by way of the humerus in the wildtype (WT) or the forelimb on the Prx1-Cre; Smad4f/f littermate embryo (PS4) at P0. Red arrow: vestigial cartilage.Lim et al.PageAuthor Manuscript Author ManuscriptFigure two. Smad4 deletion abolishes mesenchymal condensation and increases apoptosis(A, B) H E or PNA staining of sagittal sections via wild variety (WT) or Prx1-Cre; Smad4f/f (PS4) forelimb buds at E10.5 (A) or E11.five (B). (C) Representative images (left) and quantification (proper) for BrdU staining of paraffin-embedded sagittal sections of forelimbs at E11.5. BrdU signal in brown. (D) Representative images (left) and quantification (ideal) for TUNEL staining of frozen sections of forelimbs at E11.5. Apoptotic signals in green. N=3. *p0.05. Boxes denote locations for quantification.Author Manuscript Author ManuscriptDev Biol. Author manuscript; readily available in PMC 2016 April 01.Lim et al.PageAuthor Manuscript Author Manuscript Author ManuscriptFigure 3. Smad4-deficient limb bud mesenchymal cells fail to undergo condensation in micromass cultures(A) PNA or alcian blue staining of wild sort (WT) or Smad4-deficient (PS4) cultures at two, three or five days following plating. Insets Bcl-xL Inhibitor manufacturer showing higher magnification of a representative alcian bluepositive nodule present in WT but not PS4 cultures. (B) Direct fluorescence pictures of micromass cultures from mixed wild form (WT, red) and Smad4-deficient (PS4, green) cells, or Smad4-deficient (PS4, green) cells alone, at 6 days post plating. Single-channel photos for RFP or GFP shown at grey scale towards the suitable of color overlay pictures.Author ManuscriptDev Biol. Author manuscript; out there in PMC 2016 April 01.
