rb anti sox2  (Abcam)

 
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    Name:
    Anti SOX2 antibody SP76
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    ab93689
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    Structured Review

    Abcam rb anti sox2

    https://www.bioz.com/result/rb anti sox2/product/Abcam
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rb anti sox2 - by Bioz Stars, 2020-07
    99/100 stars

    Images

    Related Articles

    Staining:

    Article Title: Establishment and Characterization of a Human Small Cell Osteosarcoma Cancer Stem Cell Line: A New Possible In Vitro Model for Discovering Small Cell Osteosarcoma Biology
    Article Snippet: .. After that cells were washed three times in DPBS and stained with the mouse primary antibodies (1 : 5; anti-CD44 (Invitrogen); 1 : 5; anti-CD45 (Abcam); 1 : 5; anti-CD105 (Invitrogen); 1 : 5; anti-POU5F1 (Sigma); 1 : 10; anti-PROM1 (Miltenyi)) and with the rabbit primary antibodies (1 : 5, anti-Nanog (Abcam); 1 : 5; anti-SOX2 (Abcam); 1 : 5; anti-LIN28A (Abcam); 1 : 5; anti-KLF4 (Abcam); 1 : 5; anti-CD117 (Bioss)). .. After incubation in a humid environment at 4°C over night the primary antibodies were removed and cells were stained with the secondary antibody (1 : 300; goat anti-Mouse Alexa Fluor 635 IgG (H+L), Life Technologies; 1 : 300; goat anti-rabbit IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor 488, Invitrogen) in the dark in a humid environment at room temperature for 45 minutes.

    Incubation:

    Article Title: Sox2 Is Essential for Formation of Trophectoderm in the Preimplantation Embryo
    Article Snippet: .. Samples were transferred to a PVDF membrane (Amersham Biosciences), which was blocked in 5% w/v dried milk powder (Marvel) in 0.1% PBS-Tween, incubated with Sox2 antibody (1∶1000, Abcam) in 5% w/v dried milk powder (Marvel) at 4°C overnight and then with a peroxidase-labelled anti-rabbit IgG (1∶3000, DAKO) in 5% w/v dried milk powder (Marvel) for 1 hour. .. The enhanced chemiluminescence (ECL) detection system (Amersham Biosciences) was used to visualize the protein bands.

    Article Title: Expression of Pluripotent Stem Cell Reprogramming Factors by Prostate Tumor Initiating Cells
    Article Snippet: .. Cells were blocked with 10% goat serum/0.05% Triton X-100/phosphate buffered saline before incubation with primary antibodies, including anti-E-cadherin, anti-OCT3/4 (Santa Cruz Biotechnology), anti-SOX2 (Abcam®) and anti-β-catenin (BD Bio-sciences), overnight at 4C. .. Slides were washed, incubated with Alexa Fluor® 594 and/or 488 conjugated secondary antibodies and mounted using Vectashield® containing DAPI to counterstain nuclei.

    Article Title: Antigen presentation of the Oct4 and Sox2 peptides by CD154-activated B lymphocytes enhances the killing effect of cytotoxic T lymphocytes on tumor stem-like cells derived from cisplatin-resistant lung cancer cells
    Article Snippet: .. The slides were incubated overnight at 4o C with 1:20 rabbit anti-human Oct4 or Sox2 antibody (Abcam, USA). .. After washing with PBS the slides were incubated either with 1:800 Alexa Fluor 647 (for Sox2) or FITC (for Oct4) labeled goat anti-rabbit antibody at 37o C for 30 min followed by treatment with DAPI.

    Article Title: Progesterone Receptor Membrane Component 1 suppresses the p53 and Wnt/β-catenin pathways to promote human pluripotent stem cell self-renewal
    Article Snippet: .. To detect intracellular antigens, cells were permeabilized with 0.1% Triton X-100 after fixation with 4% PFA, and incubated with anti-OCT4 (Abcam, Cambridge, UK), anti-SOX2 (Abcam), anti-NANOG (Santa Cruz Biotechnology), anti-LC3B (Novus Biologicals), 4A68, 108-B6, and/or anti-PGRMC1 (GeneTex). .. Cells were then incubated with Alexa 488-conjugated anti-rabbit IgG (Thermo Fischer Scientific) and/or Dylight 649-conjugated anti-mouse IgG (Vector Laboratories).

    Western Blot:

    Article Title: AFF4 promotes tumorigenesis and tumor-initiation capacity of head and neck squamous cell carcinoma cells by regulating SOX2
    Article Snippet: .. The following antibodies for western blot were used in this study: anti-AFF4 (Abcam, 1:1000), anti-SOX2 (Abcam, 1:2000), anti-HA-tag (Sigma–Aldrich, 1:2000), anti-α-tubulin (Sigma–Aldrich, 1:5000). ..

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    Abcam anti sox2 rabbit monoclonal antibody solution
    <t>SOX2</t> (sex-determining region-Y homeobox-2) staining and quantification. Representative images of ( A ) SOX2 low and ( B ) high immunostaining and ( C ) digital analysis of the SOX2 high sample using StrataQuest software. Scale bars 100 μm.
    Anti Sox2 Rabbit Monoclonal Antibody Solution, supplied by Abcam, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti sox2 rabbit monoclonal antibody solution/product/Abcam
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti sox2 rabbit monoclonal antibody solution - by Bioz Stars, 2020-07
    84/100 stars
      Buy from Supplier

    96
    Abcam anti sox2 rabbit monoclonal antibody
    Phenotype of maxillary incisor germ in the F 2 generation at embryonic day 15. ( A , B ) Incisor germs of Cebpb +/+ Runx2 +/+ and Cebpb −/− Runx2 +/+ mice. Scale bar: 100 μm, 400×. Left indicates the labial side. ( C , D ) Immunostaining for SRY (sex determining region Y)-box 2 ( <t>Sox2</t> ) in incisor germs of Cebpb +/+ Runx2 +/− and Cebpb −/− Runx2 +/− mice. Scale bar: 100 μm, 400×. Left indicates labial side. Runx2 +/− tooth germ displays lingual budding(arrowhead). ( E ) Penetrance of budding according to genotype. ( F ) Comparison between Cebpb +/+ Runx2 +/− and Cebpb −/− Runx2 +/− mice lingual budding. The Y-axis on the left indicates the cell number of buddings in the maximal area of all sections; the X-axis on the bottom indicates genotypes.
    Anti Sox2 Rabbit Monoclonal Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 96/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti sox2 rabbit monoclonal antibody/product/Abcam
    Average 96 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    anti sox2 rabbit monoclonal antibody - by Bioz Stars, 2020-07
    96/100 stars
      Buy from Supplier

    99
    Abcam rb anti sox2 antibody
    Phenotype of maxillary incisor germ in the F 2 generation at embryonic day 15. ( A , B ) Incisor germs of Cebpb +/+ Runx2 +/+ and Cebpb −/− Runx2 +/+ mice. Scale bar: 100 μm, 400×. Left indicates the labial side. ( C , D ) Immunostaining for SRY (sex determining region Y)-box 2 ( <t>Sox2</t> ) in incisor germs of Cebpb +/+ Runx2 +/− and Cebpb −/− Runx2 +/− mice. Scale bar: 100 μm, 400×. Left indicates labial side. Runx2 +/− tooth germ displays lingual budding(arrowhead). ( E ) Penetrance of budding according to genotype. ( F ) Comparison between Cebpb +/+ Runx2 +/− and Cebpb −/− Runx2 +/− mice lingual budding. The Y-axis on the left indicates the cell number of buddings in the maximal area of all sections; the X-axis on the bottom indicates genotypes.
    Rb Anti Sox2 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rb anti sox2 antibody/product/Abcam
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rb anti sox2 antibody - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    Image Search Results


    SOX2 (sex-determining region-Y homeobox-2) staining and quantification. Representative images of ( A ) SOX2 low and ( B ) high immunostaining and ( C ) digital analysis of the SOX2 high sample using StrataQuest software. Scale bars 100 μm.

    Journal: Cancers

    Article Title: The Prognostic and Predictive Value of SOX2+ Cell Densities in Patients Treated for Colorectal Cancer

    doi: 10.3390/cancers12051110

    Figure Lengend Snippet: SOX2 (sex-determining region-Y homeobox-2) staining and quantification. Representative images of ( A ) SOX2 low and ( B ) high immunostaining and ( C ) digital analysis of the SOX2 high sample using StrataQuest software. Scale bars 100 μm.

    Article Snippet: Sections were then incubated for 1 h at room temperature with an anti-SOX2 rabbit monoclonal antibody solution (1:50, EPR3131, Abcam, Cambridge, UK) diluted in antibody diluent (DAKO, Denmark).

    Techniques: Staining, Immunostaining, Software

    Effect of SOX2 density on survival benefit from chemotherapy in patients with stage III CRC. Patients with stage III disease that had ( A ) low or ( B ) high SOX2 density both demonstrated significant benefit in overall survival from adjuvant chemotherapy. Patients that had ( C ) low SOX2 density did not demonstrate a benefit in cancer-specific survival whereas those with ( D ) high SOX2 density did benefit from chemotherapy. Log-rank p -values shown.

    Journal: Cancers

    Article Title: The Prognostic and Predictive Value of SOX2+ Cell Densities in Patients Treated for Colorectal Cancer

    doi: 10.3390/cancers12051110

    Figure Lengend Snippet: Effect of SOX2 density on survival benefit from chemotherapy in patients with stage III CRC. Patients with stage III disease that had ( A ) low or ( B ) high SOX2 density both demonstrated significant benefit in overall survival from adjuvant chemotherapy. Patients that had ( C ) low SOX2 density did not demonstrate a benefit in cancer-specific survival whereas those with ( D ) high SOX2 density did benefit from chemotherapy. Log-rank p -values shown.

    Article Snippet: Sections were then incubated for 1 h at room temperature with an anti-SOX2 rabbit monoclonal antibody solution (1:50, EPR3131, Abcam, Cambridge, UK) diluted in antibody diluent (DAKO, Denmark).

    Techniques:

    Prognostic value of SOX2 + cell densities. Overall survival ( A , B ) and cancer-specific survival ( C , D ) for patients with (a,c) stage II and (b,d) stage III CRC based on SOX2 expression. No significant associations between SOX2 density and survival were found. Log-rank p -values shown.

    Journal: Cancers

    Article Title: The Prognostic and Predictive Value of SOX2+ Cell Densities in Patients Treated for Colorectal Cancer

    doi: 10.3390/cancers12051110

    Figure Lengend Snippet: Prognostic value of SOX2 + cell densities. Overall survival ( A , B ) and cancer-specific survival ( C , D ) for patients with (a,c) stage II and (b,d) stage III CRC based on SOX2 expression. No significant associations between SOX2 density and survival were found. Log-rank p -values shown.

    Article Snippet: Sections were then incubated for 1 h at room temperature with an anti-SOX2 rabbit monoclonal antibody solution (1:50, EPR3131, Abcam, Cambridge, UK) diluted in antibody diluent (DAKO, Denmark).

    Techniques: Expressing

    Phenotype of maxillary incisor germ in the F 2 generation at embryonic day 15. ( A , B ) Incisor germs of Cebpb +/+ Runx2 +/+ and Cebpb −/− Runx2 +/+ mice. Scale bar: 100 μm, 400×. Left indicates the labial side. ( C , D ) Immunostaining for SRY (sex determining region Y)-box 2 ( Sox2 ) in incisor germs of Cebpb +/+ Runx2 +/− and Cebpb −/− Runx2 +/− mice. Scale bar: 100 μm, 400×. Left indicates labial side. Runx2 +/− tooth germ displays lingual budding(arrowhead). ( E ) Penetrance of budding according to genotype. ( F ) Comparison between Cebpb +/+ Runx2 +/− and Cebpb −/− Runx2 +/− mice lingual budding. The Y-axis on the left indicates the cell number of buddings in the maximal area of all sections; the X-axis on the bottom indicates genotypes.

    Journal: Scientific Reports

    Article Title: Loss of Stemness, EMT, and Supernumerary Tooth Formation in Cebpb−/−Runx2+/− Murine Incisors

    doi: 10.1038/s41598-018-23515-y

    Figure Lengend Snippet: Phenotype of maxillary incisor germ in the F 2 generation at embryonic day 15. ( A , B ) Incisor germs of Cebpb +/+ Runx2 +/+ and Cebpb −/− Runx2 +/+ mice. Scale bar: 100 μm, 400×. Left indicates the labial side. ( C , D ) Immunostaining for SRY (sex determining region Y)-box 2 ( Sox2 ) in incisor germs of Cebpb +/+ Runx2 +/− and Cebpb −/− Runx2 +/− mice. Scale bar: 100 μm, 400×. Left indicates labial side. Runx2 +/− tooth germ displays lingual budding(arrowhead). ( E ) Penetrance of budding according to genotype. ( F ) Comparison between Cebpb +/+ Runx2 +/− and Cebpb −/− Runx2 +/− mice lingual budding. The Y-axis on the left indicates the cell number of buddings in the maximal area of all sections; the X-axis on the bottom indicates genotypes.

    Article Snippet: Immunohistochemistry Heads of adult mice were decalcified with Kalkitox (Wako, Osaka, Japan) overnight at 4 °C and neutralized with 5% sodium sulfate solution (Wako) for more than 2 h. Paraffin-embedded sections were subjected to immunostaining with primary anti-Sox2 rabbit monoclonal antibody (1:100) (ab92494; Abcam, Cambridge, UK), and secondary anti-rabbit antibody (414341; Nichirei Bioscience, Tokyo, Japan), and were counterstained with hematoxylin (Wako).

    Techniques: Mouse Assay, Immunostaining

    Schematic representation of the role played by Cebpb and Runx2 in odontogenic epithelial stem cells (OESCs). Cebpb acts upstream of SRY (sex determining region Y)-box 2 ( Sox2 ). In cooperation with Oct 3/4 protein ( Pou5f1 ), Sox2 inhibits genes related to differentiation and maintains stemness in OESCs. Cebpb and Runx2 prevent epithelial-mesenchymal transition (EMT) of OESCs via snail family zinc finger 2 ( Snai2 ). Both also inhibit biglycan ( Bgn ), decorin ( Dcn ) and Bmp4 . EMT promotes expression of Sox2 and thus stemness of OESCs. EMT enables disengaged OESCs to develop supernumerary teeth by interaction with various mesenchymal stem cells such as dental MSCs.

    Journal: Scientific Reports

    Article Title: Loss of Stemness, EMT, and Supernumerary Tooth Formation in Cebpb−/−Runx2+/− Murine Incisors

    doi: 10.1038/s41598-018-23515-y

    Figure Lengend Snippet: Schematic representation of the role played by Cebpb and Runx2 in odontogenic epithelial stem cells (OESCs). Cebpb acts upstream of SRY (sex determining region Y)-box 2 ( Sox2 ). In cooperation with Oct 3/4 protein ( Pou5f1 ), Sox2 inhibits genes related to differentiation and maintains stemness in OESCs. Cebpb and Runx2 prevent epithelial-mesenchymal transition (EMT) of OESCs via snail family zinc finger 2 ( Snai2 ). Both also inhibit biglycan ( Bgn ), decorin ( Dcn ) and Bmp4 . EMT promotes expression of Sox2 and thus stemness of OESCs. EMT enables disengaged OESCs to develop supernumerary teeth by interaction with various mesenchymal stem cells such as dental MSCs.

    Article Snippet: Immunohistochemistry Heads of adult mice were decalcified with Kalkitox (Wako, Osaka, Japan) overnight at 4 °C and neutralized with 5% sodium sulfate solution (Wako) for more than 2 h. Paraffin-embedded sections were subjected to immunostaining with primary anti-Sox2 rabbit monoclonal antibody (1:100) (ab92494; Abcam, Cambridge, UK), and secondary anti-rabbit antibody (414341; Nichirei Bioscience, Tokyo, Japan), and were counterstained with hematoxylin (Wako).

    Techniques: Expressing

    Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blotting results in mHAT9d cells 48 h after transfection with Cebpb and Runx2 type1 stealth siRNA. ( A ) Changes in the expression of Cebpb , members of the Runx family, and SRY (sex determining region Y)-box 2 ( Sox2 ) in mHAT9d cells 48 h after transfection with Cebpb and Runx2 type1 stealth siRNA. Cebpb knockdown (N(negative control) + 925) shows slightly reduced expression of Sox2 . Runx2 knockdown (N + 1623) shows increased expression of Sox2 and Runx1 , but decreased expression of other Runx2 isoforms and Runx3 . Concomitant Cebpb and Runx2 type1 siRNA knockdown (925 + 1623) was also performed. ( B ) Western blotting results. N indicates negative control stealth siRNA, 925 and 1623 indicate Cebpb and Runx2 type1 siRNA, respectively (final concentration 10 nM). ( C ) Changes in the expression of adhesion molecules, epithelial-mesenchymal transition (EMT) markers, and mesenchymal markers in mHAT9d cells 48 h after transfection with Cebpb and Runx2 type1 stealth siRNA. Cebpb knockdown (N(Negative control) + 925) caused increased expression of EMT markers snail family zinc finger 2 ( Snai2 ) and decreased expression of the mesenchymal marker vimentin ( Vim ) and the epithelial marker E-cadherin ( Cdh1 ). Runx2 type1 knockdown (N + 1623) caused decreased expression of adhesion molecules Cdh1 and integrin alpha 6 ( Itga6 ), decreased expression of Vim , and increased expression of Snai2 . Concomitant Cebpb and Runx2 type1 knockdown (925 + 1623) caused decreased expression of Cdh1 , Itga6 and VIm , and increased expression of mesenchymal marker N-cadherin ( Cdh2 ) and Snai2 . ( D ) Changes in biglycan ( Bgn ), decorin ( Dcn ), bone morphogenic proteins ( Bmp s) in mHAT9d cells 48 h after transfection with Cebpb and Runx2 type1 stealth siRNA. Concomitant Cebpb and Runx2 type1 siRNA knockdown (925 + 1623) shows a markedly increased expression of Bgn and Dcn (encoding two small leucine-rich peptidogycans), and increased expression of Bmp4/6 .

    Journal: Scientific Reports

    Article Title: Loss of Stemness, EMT, and Supernumerary Tooth Formation in Cebpb−/−Runx2+/− Murine Incisors

    doi: 10.1038/s41598-018-23515-y

    Figure Lengend Snippet: Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blotting results in mHAT9d cells 48 h after transfection with Cebpb and Runx2 type1 stealth siRNA. ( A ) Changes in the expression of Cebpb , members of the Runx family, and SRY (sex determining region Y)-box 2 ( Sox2 ) in mHAT9d cells 48 h after transfection with Cebpb and Runx2 type1 stealth siRNA. Cebpb knockdown (N(negative control) + 925) shows slightly reduced expression of Sox2 . Runx2 knockdown (N + 1623) shows increased expression of Sox2 and Runx1 , but decreased expression of other Runx2 isoforms and Runx3 . Concomitant Cebpb and Runx2 type1 siRNA knockdown (925 + 1623) was also performed. ( B ) Western blotting results. N indicates negative control stealth siRNA, 925 and 1623 indicate Cebpb and Runx2 type1 siRNA, respectively (final concentration 10 nM). ( C ) Changes in the expression of adhesion molecules, epithelial-mesenchymal transition (EMT) markers, and mesenchymal markers in mHAT9d cells 48 h after transfection with Cebpb and Runx2 type1 stealth siRNA. Cebpb knockdown (N(Negative control) + 925) caused increased expression of EMT markers snail family zinc finger 2 ( Snai2 ) and decreased expression of the mesenchymal marker vimentin ( Vim ) and the epithelial marker E-cadherin ( Cdh1 ). Runx2 type1 knockdown (N + 1623) caused decreased expression of adhesion molecules Cdh1 and integrin alpha 6 ( Itga6 ), decreased expression of Vim , and increased expression of Snai2 . Concomitant Cebpb and Runx2 type1 knockdown (925 + 1623) caused decreased expression of Cdh1 , Itga6 and VIm , and increased expression of mesenchymal marker N-cadherin ( Cdh2 ) and Snai2 . ( D ) Changes in biglycan ( Bgn ), decorin ( Dcn ), bone morphogenic proteins ( Bmp s) in mHAT9d cells 48 h after transfection with Cebpb and Runx2 type1 stealth siRNA. Concomitant Cebpb and Runx2 type1 siRNA knockdown (925 + 1623) shows a markedly increased expression of Bgn and Dcn (encoding two small leucine-rich peptidogycans), and increased expression of Bmp4/6 .

    Article Snippet: Immunohistochemistry Heads of adult mice were decalcified with Kalkitox (Wako, Osaka, Japan) overnight at 4 °C and neutralized with 5% sodium sulfate solution (Wako) for more than 2 h. Paraffin-embedded sections were subjected to immunostaining with primary anti-Sox2 rabbit monoclonal antibody (1:100) (ab92494; Abcam, Cambridge, UK), and secondary anti-rabbit antibody (414341; Nichirei Bioscience, Tokyo, Japan), and were counterstained with hematoxylin (Wako).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Transfection, Expressing, Negative Control, Concentration Assay, Marker

    Phenotypes of incisors in Cebpb −/− mice maintained in a 129 Sv background. ( A ) Three-dimensional image of the skull. The left incisor is short. Scale bar: 10 mm. ( B ) Micro-computed tomography (CT) axial section image at the level of the maxillary incisor. Left incisors are deformed. ( C ) Sagittal hematoxylin-eosin (H E)-stained section of the left upper incisor. Ectopic marked growth of dentin and enamel is observed in the apical region of the left incisor. Scale bar: 1 mm, 40×. ( D ) Amelogenin immunostaining (red) and Hoechst nuclear staining (blue). Amelogenin is observed in the hypocalcification site. Mature enamel was removed from this decalcified section leaving a clear enamel space (ES). Arrowheads indicate epithelial pearls. Scale bar: 100 μm, 100×. ( E ) Many dental tubules can be seen in dentin, which is also more abundant. Derangement of ameloblasts are observed near enamel matrix. Scale bar: 100 μm, 200×. ( F ) Magnified image of dental tubules in dentin. Scale bar:200 μm. 400× Inset: 1000×. ( G ) Nearly normal development of the right incisor. Ameloblasts, enamel-producing cells, are arranged in a single-cell row. Odontoblasts, dentin-producing cells, line the pulp cavity. Increased dentin is observed in both the labial and lingual side. Miniaturization of the labial cervical loop epithelium is seen. Scale bar: 1 mm, 40×. ( H ) Sagittal H E-stained section of the left mandibular incisor of a 10-month-old mouse. Scale bar: 2 mm, 20×. ( I ) The boxed region in H. A mature tooth structure with pulp enclosed with dentin in a different direction is observed in a periapical region of a mandibular incisor. Scale bar: 500 μm, 100×. ( J ) SRY (sex determining region Y)-box 2 ( Sox2 ) - Positive cells in a maxillary incisor labial cervical loop epithelium of 1-month-old wild-type (WT) mice. Scale bar: 100 μm, 400×. ( K ) Sox2-Positive cells in a maxillary incisor labial cervical loop epithelium of 19-month-old Cebpb −/− mice. Scale bar: 100 μm, 400×. ( L ) Comparison between WT and Cebpb −/− mice. Y-axis indicates the labial cervical loop epithelium area; X-axis indicates genotypes. ( M ) Comparison between WT and Cebpb −/− mice. Y-axis indicates number of Sox2-positive cells in the labial cervical loop epithelium; the X-axis indicates genotypes. ( N ) Comparison between WT and Cebpb −/− mice. Y-axis indicates Sox2-positive area in the labial cervical loop epithetium area; X-axis indicates genotypes.

    Journal: Scientific Reports

    Article Title: Loss of Stemness, EMT, and Supernumerary Tooth Formation in Cebpb−/−Runx2+/− Murine Incisors

    doi: 10.1038/s41598-018-23515-y

    Figure Lengend Snippet: Phenotypes of incisors in Cebpb −/− mice maintained in a 129 Sv background. ( A ) Three-dimensional image of the skull. The left incisor is short. Scale bar: 10 mm. ( B ) Micro-computed tomography (CT) axial section image at the level of the maxillary incisor. Left incisors are deformed. ( C ) Sagittal hematoxylin-eosin (H E)-stained section of the left upper incisor. Ectopic marked growth of dentin and enamel is observed in the apical region of the left incisor. Scale bar: 1 mm, 40×. ( D ) Amelogenin immunostaining (red) and Hoechst nuclear staining (blue). Amelogenin is observed in the hypocalcification site. Mature enamel was removed from this decalcified section leaving a clear enamel space (ES). Arrowheads indicate epithelial pearls. Scale bar: 100 μm, 100×. ( E ) Many dental tubules can be seen in dentin, which is also more abundant. Derangement of ameloblasts are observed near enamel matrix. Scale bar: 100 μm, 200×. ( F ) Magnified image of dental tubules in dentin. Scale bar:200 μm. 400× Inset: 1000×. ( G ) Nearly normal development of the right incisor. Ameloblasts, enamel-producing cells, are arranged in a single-cell row. Odontoblasts, dentin-producing cells, line the pulp cavity. Increased dentin is observed in both the labial and lingual side. Miniaturization of the labial cervical loop epithelium is seen. Scale bar: 1 mm, 40×. ( H ) Sagittal H E-stained section of the left mandibular incisor of a 10-month-old mouse. Scale bar: 2 mm, 20×. ( I ) The boxed region in H. A mature tooth structure with pulp enclosed with dentin in a different direction is observed in a periapical region of a mandibular incisor. Scale bar: 500 μm, 100×. ( J ) SRY (sex determining region Y)-box 2 ( Sox2 ) - Positive cells in a maxillary incisor labial cervical loop epithelium of 1-month-old wild-type (WT) mice. Scale bar: 100 μm, 400×. ( K ) Sox2-Positive cells in a maxillary incisor labial cervical loop epithelium of 19-month-old Cebpb −/− mice. Scale bar: 100 μm, 400×. ( L ) Comparison between WT and Cebpb −/− mice. Y-axis indicates the labial cervical loop epithelium area; X-axis indicates genotypes. ( M ) Comparison between WT and Cebpb −/− mice. Y-axis indicates number of Sox2-positive cells in the labial cervical loop epithelium; the X-axis indicates genotypes. ( N ) Comparison between WT and Cebpb −/− mice. Y-axis indicates Sox2-positive area in the labial cervical loop epithetium area; X-axis indicates genotypes.

    Article Snippet: Immunohistochemistry Heads of adult mice were decalcified with Kalkitox (Wako, Osaka, Japan) overnight at 4 °C and neutralized with 5% sodium sulfate solution (Wako) for more than 2 h. Paraffin-embedded sections were subjected to immunostaining with primary anti-Sox2 rabbit monoclonal antibody (1:100) (ab92494; Abcam, Cambridge, UK), and secondary anti-rabbit antibody (414341; Nichirei Bioscience, Tokyo, Japan), and were counterstained with hematoxylin (Wako).

    Techniques: Mouse Assay, Micro-CT, Staining, Immunostaining

    Phenotype of maxillary incisor germ in the F 2 generation at embryonic day 15. ( A , B ) Incisor germs of Cebpb +/+ Runx2 +/+ and Cebpb −/− Runx2 +/+ mice. Scale bar: 100 μm, 400×. Left indicates the labial side. ( C , D ) Immunostaining for SRY (sex determining region Y)-box 2 ( Sox2 ) in incisor germs of Cebpb +/+ Runx2 +/− and Cebpb −/− Runx2 +/− mice. Scale bar: 100 μm, 400×. Left indicates labial side. Runx2 +/− tooth germ displays lingual budding(arrowhead). ( E ) Penetrance of budding according to genotype. ( F ) Comparison between Cebpb +/+ Runx2 +/− and Cebpb −/− Runx2 +/− mice lingual budding. The Y-axis on the left indicates the cell number of buddings in the maximal area of all sections; the X-axis on the bottom indicates genotypes.

    Journal: Scientific Reports

    Article Title: Loss of Stemness, EMT, and Supernumerary Tooth Formation in Cebpb−/−Runx2+/− Murine Incisors

    doi: 10.1038/s41598-018-23515-y

    Figure Lengend Snippet: Phenotype of maxillary incisor germ in the F 2 generation at embryonic day 15. ( A , B ) Incisor germs of Cebpb +/+ Runx2 +/+ and Cebpb −/− Runx2 +/+ mice. Scale bar: 100 μm, 400×. Left indicates the labial side. ( C , D ) Immunostaining for SRY (sex determining region Y)-box 2 ( Sox2 ) in incisor germs of Cebpb +/+ Runx2 +/− and Cebpb −/− Runx2 +/− mice. Scale bar: 100 μm, 400×. Left indicates labial side. Runx2 +/− tooth germ displays lingual budding(arrowhead). ( E ) Penetrance of budding according to genotype. ( F ) Comparison between Cebpb +/+ Runx2 +/− and Cebpb −/− Runx2 +/− mice lingual budding. The Y-axis on the left indicates the cell number of buddings in the maximal area of all sections; the X-axis on the bottom indicates genotypes.

    Article Snippet: Heads of adult mice were decalcified with Kalkitox (Wako, Osaka, Japan) overnight at 4 °C and neutralized with 5% sodium sulfate solution (Wako) for more than 2 h. Paraffin-embedded sections were subjected to immunostaining with primary anti-Sox2 rabbit monoclonal antibody (1:100) (ab92494; Abcam, Cambridge, UK), and secondary anti-rabbit antibody (414341; Nichirei Bioscience, Tokyo, Japan), and were counterstained with hematoxylin (Wako).

    Techniques: Mouse Assay, Immunostaining

    Schematic representation of the role played by Cebpb and Runx2 in odontogenic epithelial stem cells (OESCs). Cebpb acts upstream of SRY (sex determining region Y)-box 2 ( Sox2 ). In cooperation with Oct 3/4 protein ( Pou5f1 ), Sox2 inhibits genes related to differentiation and maintains stemness in OESCs. Cebpb and Runx2 prevent epithelial-mesenchymal transition (EMT) of OESCs via snail family zinc finger 2 ( Snai2 ). Both also inhibit biglycan ( Bgn ), decorin ( Dcn ) and Bmp4 . EMT promotes expression of Sox2 and thus stemness of OESCs. EMT enables disengaged OESCs to develop supernumerary teeth by interaction with various mesenchymal stem cells such as dental MSCs.

    Journal: Scientific Reports

    Article Title: Loss of Stemness, EMT, and Supernumerary Tooth Formation in Cebpb−/−Runx2+/− Murine Incisors

    doi: 10.1038/s41598-018-23515-y

    Figure Lengend Snippet: Schematic representation of the role played by Cebpb and Runx2 in odontogenic epithelial stem cells (OESCs). Cebpb acts upstream of SRY (sex determining region Y)-box 2 ( Sox2 ). In cooperation with Oct 3/4 protein ( Pou5f1 ), Sox2 inhibits genes related to differentiation and maintains stemness in OESCs. Cebpb and Runx2 prevent epithelial-mesenchymal transition (EMT) of OESCs via snail family zinc finger 2 ( Snai2 ). Both also inhibit biglycan ( Bgn ), decorin ( Dcn ) and Bmp4 . EMT promotes expression of Sox2 and thus stemness of OESCs. EMT enables disengaged OESCs to develop supernumerary teeth by interaction with various mesenchymal stem cells such as dental MSCs.

    Article Snippet: Heads of adult mice were decalcified with Kalkitox (Wako, Osaka, Japan) overnight at 4 °C and neutralized with 5% sodium sulfate solution (Wako) for more than 2 h. Paraffin-embedded sections were subjected to immunostaining with primary anti-Sox2 rabbit monoclonal antibody (1:100) (ab92494; Abcam, Cambridge, UK), and secondary anti-rabbit antibody (414341; Nichirei Bioscience, Tokyo, Japan), and were counterstained with hematoxylin (Wako).

    Techniques: Expressing

    Phenotypes of incisors in Cebpb −/− mice maintained in a 129 Sv background. ( A ) Three-dimensional image of the skull. The left incisor is short. Scale bar: 10 mm. ( B ) Micro-computed tomography (CT) axial section image at the level of the maxillary incisor. Left incisors are deformed. ( C ) Sagittal hematoxylin-eosin (H E)-stained section of the left upper incisor. Ectopic marked growth of dentin and enamel is observed in the apical region of the left incisor. Scale bar: 1 mm, 40×. ( D ) Amelogenin immunostaining (red) and Hoechst nuclear staining (blue). Amelogenin is observed in the hypocalcification site. Mature enamel was removed from this decalcified section leaving a clear enamel space (ES). Arrowheads indicate epithelial pearls. Scale bar: 100 μm, 100×. ( E ) Many dental tubules can be seen in dentin, which is also more abundant. Derangement of ameloblasts are observed near enamel matrix. Scale bar: 100 μm, 200×. ( F ) Magnified image of dental tubules in dentin. Scale bar:200 μm. 400× Inset: 1000×. ( G ) Nearly normal development of the right incisor. Ameloblasts, enamel-producing cells, are arranged in a single-cell row. Odontoblasts, dentin-producing cells, line the pulp cavity. Increased dentin is observed in both the labial and lingual side. Miniaturization of the labial cervical loop epithelium is seen. Scale bar: 1 mm, 40×. ( H ) Sagittal H E-stained section of the left mandibular incisor of a 10-month-old mouse. Scale bar: 2 mm, 20×. ( I ) The boxed region in H. A mature tooth structure with pulp enclosed with dentin in a different direction is observed in a periapical region of a mandibular incisor. Scale bar: 500 μm, 100×. ( J ) SRY (sex determining region Y)-box 2 ( Sox2 ) - Positive cells in a maxillary incisor labial cervical loop epithelium of 1-month-old wild-type (WT) mice. Scale bar: 100 μm, 400×. ( K ) Sox2-Positive cells in a maxillary incisor labial cervical loop epithelium of 19-month-old Cebpb −/− mice. Scale bar: 100 μm, 400×. ( L ) Comparison between WT and Cebpb −/− mice. Y-axis indicates the labial cervical loop epithelium area; X-axis indicates genotypes. ( M ) Comparison between WT and Cebpb −/− mice. Y-axis indicates number of Sox2-positive cells in the labial cervical loop epithelium; the X-axis indicates genotypes. ( N ) Comparison between WT and Cebpb −/− mice. Y-axis indicates Sox2-positive area in the labial cervical loop epithetium area; X-axis indicates genotypes.

    Journal: Scientific Reports

    Article Title: Loss of Stemness, EMT, and Supernumerary Tooth Formation in Cebpb−/−Runx2+/− Murine Incisors

    doi: 10.1038/s41598-018-23515-y

    Figure Lengend Snippet: Phenotypes of incisors in Cebpb −/− mice maintained in a 129 Sv background. ( A ) Three-dimensional image of the skull. The left incisor is short. Scale bar: 10 mm. ( B ) Micro-computed tomography (CT) axial section image at the level of the maxillary incisor. Left incisors are deformed. ( C ) Sagittal hematoxylin-eosin (H E)-stained section of the left upper incisor. Ectopic marked growth of dentin and enamel is observed in the apical region of the left incisor. Scale bar: 1 mm, 40×. ( D ) Amelogenin immunostaining (red) and Hoechst nuclear staining (blue). Amelogenin is observed in the hypocalcification site. Mature enamel was removed from this decalcified section leaving a clear enamel space (ES). Arrowheads indicate epithelial pearls. Scale bar: 100 μm, 100×. ( E ) Many dental tubules can be seen in dentin, which is also more abundant. Derangement of ameloblasts are observed near enamel matrix. Scale bar: 100 μm, 200×. ( F ) Magnified image of dental tubules in dentin. Scale bar:200 μm. 400× Inset: 1000×. ( G ) Nearly normal development of the right incisor. Ameloblasts, enamel-producing cells, are arranged in a single-cell row. Odontoblasts, dentin-producing cells, line the pulp cavity. Increased dentin is observed in both the labial and lingual side. Miniaturization of the labial cervical loop epithelium is seen. Scale bar: 1 mm, 40×. ( H ) Sagittal H E-stained section of the left mandibular incisor of a 10-month-old mouse. Scale bar: 2 mm, 20×. ( I ) The boxed region in H. A mature tooth structure with pulp enclosed with dentin in a different direction is observed in a periapical region of a mandibular incisor. Scale bar: 500 μm, 100×. ( J ) SRY (sex determining region Y)-box 2 ( Sox2 ) - Positive cells in a maxillary incisor labial cervical loop epithelium of 1-month-old wild-type (WT) mice. Scale bar: 100 μm, 400×. ( K ) Sox2-Positive cells in a maxillary incisor labial cervical loop epithelium of 19-month-old Cebpb −/− mice. Scale bar: 100 μm, 400×. ( L ) Comparison between WT and Cebpb −/− mice. Y-axis indicates the labial cervical loop epithelium area; X-axis indicates genotypes. ( M ) Comparison between WT and Cebpb −/− mice. Y-axis indicates number of Sox2-positive cells in the labial cervical loop epithelium; the X-axis indicates genotypes. ( N ) Comparison between WT and Cebpb −/− mice. Y-axis indicates Sox2-positive area in the labial cervical loop epithetium area; X-axis indicates genotypes.

    Article Snippet: Heads of adult mice were decalcified with Kalkitox (Wako, Osaka, Japan) overnight at 4 °C and neutralized with 5% sodium sulfate solution (Wako) for more than 2 h. Paraffin-embedded sections were subjected to immunostaining with primary anti-Sox2 rabbit monoclonal antibody (1:100) (ab92494; Abcam, Cambridge, UK), and secondary anti-rabbit antibody (414341; Nichirei Bioscience, Tokyo, Japan), and were counterstained with hematoxylin (Wako).

    Techniques: Mouse Assay, Micro-CT, Staining, Immunostaining

    Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blotting results in mHAT9d cells 48 h after transfection with Cebpb and Runx2 type1 stealth siRNA. ( A ) Changes in the expression of Cebpb , members of the Runx family, and SRY (sex determining region Y)-box 2 ( Sox2 ) in mHAT9d cells 48 h after transfection with Cebpb and Runx2 type1 stealth siRNA. Cebpb knockdown (N(negative control) + 925) shows slightly reduced expression of Sox2 . Runx2 knockdown (N + 1623) shows increased expression of Sox2 and Runx1 , but decreased expression of other Runx2 isoforms and Runx3 . Concomitant Cebpb and Runx2 type1 siRNA knockdown (925 + 1623) was also performed. ( B ) Western blotting results. N indicates negative control stealth siRNA, 925 and 1623 indicate Cebpb and Runx2 type1 siRNA, respectively (final concentration 10 nM). ( C ) Changes in the expression of adhesion molecules, epithelial-mesenchymal transition (EMT) markers, and mesenchymal markers in mHAT9d cells 48 h after transfection with Cebpb and Runx2 type1 stealth siRNA. Cebpb knockdown (N(Negative control) + 925) caused increased expression of EMT markers snail family zinc finger 2 ( Snai2 ) and decreased expression of the mesenchymal marker vimentin ( Vim ) and the epithelial marker E-cadherin ( Cdh1 ). Runx2 type1 knockdown (N + 1623) caused decreased expression of adhesion molecules Cdh1 and integrin alpha 6 ( Itga6 ), decreased expression of Vim , and increased expression of Snai2 . Concomitant Cebpb and Runx2 type1 knockdown (925 + 1623) caused decreased expression of Cdh1 , Itga6 and VIm , and increased expression of mesenchymal marker N-cadherin ( Cdh2 ) and Snai2 . ( D ) Changes in biglycan ( Bgn ), decorin ( Dcn ), bone morphogenic proteins ( Bmp s) in mHAT9d cells 48 h after transfection with Cebpb and Runx2 type1 stealth siRNA. Concomitant Cebpb and Runx2 type1 siRNA knockdown (925 + 1623) shows a markedly increased expression of Bgn and Dcn (encoding two small leucine-rich peptidogycans), and increased expression of Bmp4/6 .

    Journal: Scientific Reports

    Article Title: Loss of Stemness, EMT, and Supernumerary Tooth Formation in Cebpb−/−Runx2+/− Murine Incisors

    doi: 10.1038/s41598-018-23515-y

    Figure Lengend Snippet: Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blotting results in mHAT9d cells 48 h after transfection with Cebpb and Runx2 type1 stealth siRNA. ( A ) Changes in the expression of Cebpb , members of the Runx family, and SRY (sex determining region Y)-box 2 ( Sox2 ) in mHAT9d cells 48 h after transfection with Cebpb and Runx2 type1 stealth siRNA. Cebpb knockdown (N(negative control) + 925) shows slightly reduced expression of Sox2 . Runx2 knockdown (N + 1623) shows increased expression of Sox2 and Runx1 , but decreased expression of other Runx2 isoforms and Runx3 . Concomitant Cebpb and Runx2 type1 siRNA knockdown (925 + 1623) was also performed. ( B ) Western blotting results. N indicates negative control stealth siRNA, 925 and 1623 indicate Cebpb and Runx2 type1 siRNA, respectively (final concentration 10 nM). ( C ) Changes in the expression of adhesion molecules, epithelial-mesenchymal transition (EMT) markers, and mesenchymal markers in mHAT9d cells 48 h after transfection with Cebpb and Runx2 type1 stealth siRNA. Cebpb knockdown (N(Negative control) + 925) caused increased expression of EMT markers snail family zinc finger 2 ( Snai2 ) and decreased expression of the mesenchymal marker vimentin ( Vim ) and the epithelial marker E-cadherin ( Cdh1 ). Runx2 type1 knockdown (N + 1623) caused decreased expression of adhesion molecules Cdh1 and integrin alpha 6 ( Itga6 ), decreased expression of Vim , and increased expression of Snai2 . Concomitant Cebpb and Runx2 type1 knockdown (925 + 1623) caused decreased expression of Cdh1 , Itga6 and VIm , and increased expression of mesenchymal marker N-cadherin ( Cdh2 ) and Snai2 . ( D ) Changes in biglycan ( Bgn ), decorin ( Dcn ), bone morphogenic proteins ( Bmp s) in mHAT9d cells 48 h after transfection with Cebpb and Runx2 type1 stealth siRNA. Concomitant Cebpb and Runx2 type1 siRNA knockdown (925 + 1623) shows a markedly increased expression of Bgn and Dcn (encoding two small leucine-rich peptidogycans), and increased expression of Bmp4/6 .

    Article Snippet: Heads of adult mice were decalcified with Kalkitox (Wako, Osaka, Japan) overnight at 4 °C and neutralized with 5% sodium sulfate solution (Wako) for more than 2 h. Paraffin-embedded sections were subjected to immunostaining with primary anti-Sox2 rabbit monoclonal antibody (1:100) (ab92494; Abcam, Cambridge, UK), and secondary anti-rabbit antibody (414341; Nichirei Bioscience, Tokyo, Japan), and were counterstained with hematoxylin (Wako).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Transfection, Expressing, Negative Control, Concentration Assay, Marker