Journal: The American journal of pathology

Article Title: Loss of β-catenin induces multifocal periosteal chondroma-like masses in mice.

doi: 10.1016/j.ajpath.2012.11.012

Figure Lengend Snippet: Figure 2 Outgrowth of growth plate and ectopic cartilage in perios- teum in tamoxifen-injected Col2CreER;b-catenin fl/fl mice. Col2CreER; b-catenin fl/flmice (CeJ) and their littermate b-cateninfl/flmice (A and B) received 100 mg/10 mL i.p. injections of tamoxifen per mouse at P5 to P7. Longitudinal sections of knee joints were prepared at 2 weeks (A, C, E, G, and I) or 5 weeks (B, D, F, H, and J) after completion of the tamoxifen injections. Arrows indicate out-growing growth plate (C) or ectopic cartilage masses embedded in the periosteum (D, H, and J). Boxed areas in C and D are enlarged in E and F, respectively.

Article Snippet: The American Journal of Pathology - ajp.amjpathol.org The mouse growth plate paraffin sections were incubated with the rabbit polyclonal antieb-catenin antibody (1:250; Cell Signaling Technology, Inc., Danvers, MA), followed by incubation with horseradish peroxidase conjugated antirabbit antibody (1:100; Vector Laboratories) and ImmPACT SG peroxidase substrate (Vector Laboratories) with methyl green counter staining.

Techniques: Injection

Journal: The American journal of pathology

Article Title: Loss of β-catenin induces multifocal periosteal chondroma-like masses in mice.

doi: 10.1016/j.ajpath.2012.11.012

Figure Lengend Snippet: Figure 3 Localization of Cre-positive cells after tamoxifen injection in Col2CreER;b-catenin fl/fl;Rosa-LacZ mice. Col2CreER;b-catenin fl/fl;Rosa-LacZ mice (A, B, and DeF) and littermate b-catenin fl/fl;Rosa-LacZ mice (C) received 100 mg/10 mL i.p. injections of tamoxifen per mouse at P5 and P7. Longitudinal sections of knee joints were prepared at 1 (AeC), 2 (D and E), or 8 (F) weeks after completion of the tamoxifen injections and subjected to b-galactosidase staining. B and E: Magnified images of the boxed areas in A and D, respectively.

Article Snippet: The American Journal of Pathology - ajp.amjpathol.org The mouse growth plate paraffin sections were incubated with the rabbit polyclonal antieb-catenin antibody (1:250; Cell Signaling Technology, Inc., Danvers, MA), followed by incubation with horseradish peroxidase conjugated antirabbit antibody (1:100; Vector Laboratories) and ImmPACT SG peroxidase substrate (Vector Laboratories) with methyl green counter staining.

Techniques: Injection, Staining

Journal: The American journal of pathology

Article Title: Loss of β-catenin induces multifocal periosteal chondroma-like masses in mice.

doi: 10.1016/j.ajpath.2012.11.012

Figure Lengend Snippet: Figure 4 Imbalance of cell proliferating activity in growth plate in the tamoxifen injected Col2CreER;b-catenin fl/flmice. Col2CreER;b-catenin fl/fl;Rosa- LacZ mice (A and B) and littermate b-cateninfl/fl;Rosa-LacZ mice (C and D) received 100 mg/10 mL i.p. injections of tamoxifen per mouse at P5 and P7. Longitudinal sections of knee joints were prepared at 2 weeks after completion of the tamoxifen injections and subjected to PCNA staining. E: Ratios of PCNA- positive cells were measured (see Materials and Methods). *P < 0.05

Article Snippet: The American Journal of Pathology - ajp.amjpathol.org The mouse growth plate paraffin sections were incubated with the rabbit polyclonal antieb-catenin antibody (1:250; Cell Signaling Technology, Inc., Danvers, MA), followed by incubation with horseradish peroxidase conjugated antirabbit antibody (1:100; Vector Laboratories) and ImmPACT SG peroxidase substrate (Vector Laboratories) with methyl green counter staining.

Techniques: Activity Assay, Injection, Staining

Journal: The American journal of pathology

Article Title: Loss of β-catenin induces multifocal periosteal chondroma-like masses in mice.

doi: 10.1016/j.ajpath.2012.11.012

Figure Lengend Snippet: Figure 5 PCNA, collagen 10, TUNEL, and TRAP staining in ectopic cartilaginous masses at early and late stages after tamoxifen injections. Col2CreER;b-catenin fl/fl;Rosa-LacZ mice received 100 mg/10 mL i.p. injec- tions of tamoxifen per mouse at P5 and P7. Longitudinal sections of knee joints were prepared at 2 weeks (A, C, E, G, and I) or 8 weeks (B, D, F, H, and J) after completion of the tamoxifen injections and subjected to PCNA (A and B), collagen 10 immunostaining (C and D), TUNEL (E and F), and TRAP (I and J) staining. The arrows in E indicates apoptotic cells. G and H: Phase contrast images (E and F), respectively.

Article Snippet: The American Journal of Pathology - ajp.amjpathol.org The mouse growth plate paraffin sections were incubated with the rabbit polyclonal antieb-catenin antibody (1:250; Cell Signaling Technology, Inc., Danvers, MA), followed by incubation with horseradish peroxidase conjugated antirabbit antibody (1:100; Vector Laboratories) and ImmPACT SG peroxidase substrate (Vector Laboratories) with methyl green counter staining.

Techniques: TUNEL Assay, Staining, Immunostaining

Journal: The American journal of pathology

Article Title: Loss of β-catenin induces multifocal periosteal chondroma-like masses in mice.

doi: 10.1016/j.ajpath.2012.11.012

Figure Lengend Snippet: Figure 8 Immunolocalization of b-catenin in ostechondromas and rib growth plates. AeL: Sections of osteochondromas from HME patients (A, B, E, F, I, and J) and rib growth plate obtained from autopsy (C, D, G, H, K, and L) were subjected to immunofluorescence staining for b-catenin. Cartilage caps of osteo- chondromas were divided into two parts: a region composing of solitary and relatively small cells (Solitary), and a region adjacent to the bone and composed of relatively large cells (Large). Resting zone (Resting) and hypertrophic zone (Hypertrophy) of growth plates were separately inspected. AeD: b-catenin staining; (EeH) merged images of b-catenin staining (red) and DAPI nuclear staining (blue); (IeL) phase contrast images (AeD), respectively. M: Ratios of b-catenin positive cells were evaluated as previously described (see Materials and Methods). *P < 0.05.

Article Snippet: The American Journal of Pathology - ajp.amjpathol.org The mouse growth plate paraffin sections were incubated with the rabbit polyclonal antieb-catenin antibody (1:250; Cell Signaling Technology, Inc., Danvers, MA), followed by incubation with horseradish peroxidase conjugated antirabbit antibody (1:100; Vector Laboratories) and ImmPACT SG peroxidase substrate (Vector Laboratories) with methyl green counter staining.

Techniques: Staining

Journal: Journal of Investigative Dermatology

Article Title: Microtubule-Dependent Redistribution of a Cytoplasmic Cornified Envelope Precursor

doi: 10.1046/j.0022-202x.2003.22105.x

Figure Lengend Snippet: Figure 3 S100A11 redistribution to the cell periphery requires intact tubulin filaments. Keratinocytes, growing on coverslips, were treated for 2 h at 37C in the absence or presence of 1 mM vincristine, a microtubule destabilizer. The cells were then maintained for 1 h in medium containing 0.09 or 0.3 mM calcium, fixed, permeabilized, and costained with anti-S100A11 (green) and anti-b-tubulin (red). The images were obtained by confocal microscopy. The overlay image indicates the combination of the S100A11 and b-tubulin images. Colocalization is identified by yellow color. Circles, the perimeter of the nuclei; arrows, indicate the direction of S100A11 redistribution. No arrows are shown in the b-tubulin panel, because b-tubulin does not move in response to calcium treatment. In the vincristine-treated images, the arrows indicate the direction of tubulin retreat from the cell periphery. Bars, 10 mm.

Article Snippet: Total protein (500 mg) was incubated with 2 mg of mouse antib-tubulin antibody (sc5274, Santa Cruz Biotechnology) or 1 mg of rabbit anti-S100A11 antibody for 3 h at 41C.

Techniques: Confocal Microscopy

Journal: Journal of Investigative Dermatology

Article Title: Microtubule-Dependent Redistribution of a Cytoplasmic Cornified Envelope Precursor

doi: 10.1046/j.0022-202x.2003.22105.x

Figure Lengend Snippet: Figure 4 S100A10 does not colocalize with tubulin filaments. Human keratinocytes were cultured on coverslips and treated with low (0.09 mM) or high (0.3 mM) calcium-containing medium for 1 h at 371C. The cells were then fixed, permeabilized, and immunostained with anti- S100A10 (green) or anti-b-tubulin (red). The overlay panels indicate the combined images. Bars, 10 mm.

Article Snippet: Total protein (500 mg) was incubated with 2 mg of mouse antib-tubulin antibody (sc5274, Santa Cruz Biotechnology) or 1 mg of rabbit anti-S100A11 antibody for 3 h at 41C.

Techniques: Cell Culture

Journal: Journal of Investigative Dermatology

Article Title: Microtubule-Dependent Redistribution of a Cytoplasmic Cornified Envelope Precursor

doi: 10.1046/j.0022-202x.2003.22105.x

Figure Lengend Snippet: Figure 5 Coprecipitation of S100A11 and b-tubulin. (A) Extracts from human keratinocytes grown in 0.09 mM calcium-containing medium were prepared in lysis buffer and then precipitated with anti-b-tubulin or nonimmune serum. The precipitated samples were then electrophor- esed and blotted with anti-S100A11. Total extract (TE) was electro- phoresed in a parallel lane. Migration of the S100A11 monomer, and higher-molecular-weight immunoreactive forms () are indicated. (B) Extracts, prepared as above, were precipitated using anti-S100A11 or nonimmune serum, electrophoresed, and blotted with anti-b-tubulin. TE was electrophoresed in a parallel lane. Antibody binding was visualized using the appropriate secondary antibody and chemilumi- nescent detection reagent. (C) Cell supernatant, membrane, and particulate fractions were prepared as outlined under Materials and Methods. Equal cell equivalents of each fraction were electrophoresed and immunoblotted with anti-S100A11. The S100A11 monomer is indicated, as is the migration of the larger multimers (). No signal was observed when nonimmune serum was substituted for anti-S100A11.

Article Snippet: Total protein (500 mg) was incubated with 2 mg of mouse antib-tubulin antibody (sc5274, Santa Cruz Biotechnology) or 1 mg of rabbit anti-S100A11 antibody for 3 h at 41C.

Techniques: Lysis, Migration, Molecular Weight, Binding Assay, Membrane