Journal: The International journal of developmental biology

Article Title: Chromatin organization and timing of polar body I extrusion identify developmentally competent mouse oocytes.

doi: 10.1387/ijdb.180362sg

Figure Lengend Snippet: Fig. 4. Immuno-detection of CDX2- and OCT4-positive blastomeres in blastocysts. Blastocysts developed from fertilized group A or ovulated MII oocytes. Blastocysts were counterstained with DAPI used to count the total number of blastomeres. The number of blastomeres was not significantly different (p> 0.05) among the two groups analyzed. Bar, 10 mm.

Article Snippet: Embryos were then processed for sequential immunolabeling using a rabbit polyclonal anti-human OCT4 (Abcam, cat. n. ab19857) and a rabbit anti-human CDX2 (Cell Signaling, cat. n 3977S) antibody.

Techniques:

Journal: Molecular Medicine Reports

Article Title: Overexpression of caudal type homeobox transcription factor 2 inhibits the growth of the MGC-803 human gastric cancer cell line in vivo

doi: 10.3892/mmr.2015.3413

Figure Lengend Snippet: Sequences of the primers used for reverse transcription semi-quantitative polymerase chain reaction.

Article Snippet: Specific rabbit anti-human polyclonal antibodies to CDX2 (#12306), c-Myc (#5605), Skp2 (#2652), Bax (#5023), Bcl-2 (#2827), cyclin D1 (#2978), survivin (#2808) and GAPDH (#2118) were provided by Cell Signaling Technology, Inc. (Beverly, MA, USA) .

Techniques: Reverse Transcription, Sequencing

Journal: Molecular Medicine Reports

Article Title: Overexpression of caudal type homeobox transcription factor 2 inhibits the growth of the MGC-803 human gastric cancer cell line in vivo

doi: 10.3892/mmr.2015.3413

Figure Lengend Snippet: Determination of lentivirus titers. The viral titers of recombinant lentivirus transfected into 293T cells were determined by end point dilution, through counting the numbers of fluorescent green-labeled cells under a fluorescence microscope (magnification, x100). The viral dilution factor was 1:1,000. LV-GFP, lentivirus-green fluorescent protein; NC, negative control; CDX2, caudal type homeobox transcription factor 2.

Article Snippet: Specific rabbit anti-human polyclonal antibodies to CDX2 (#12306), c-Myc (#5605), Skp2 (#2652), Bax (#5023), Bcl-2 (#2827), cyclin D1 (#2978), survivin (#2808) and GAPDH (#2118) were provided by Cell Signaling Technology, Inc. (Beverly, MA, USA) .

Techniques: Recombinant, Transfection, Labeling, Fluorescence, Microscopy, Negative Control

Journal: Molecular Medicine Reports

Article Title: Overexpression of caudal type homeobox transcription factor 2 inhibits the growth of the MGC-803 human gastric cancer cell line in vivo

doi: 10.3892/mmr.2015.3413

Figure Lengend Snippet: Overexpression of CDX2 inhibits tumor growth and induces tumor cell apoptosis in LV-GFP-CDX2-treated mice. (A) Relative tumor volume growth curve revealed significant growth tendencies in the PBS and LV-GFP-NC groups, while the MGC-803 tumor growth in the LV-GFP-CDX2 group was markedly inhibited. (B) Tumor volumes in the LV-GFP-CDX2 groupe were smaller compared with those in the control group 14 days after tumor injection ( * P<0.05). (C) Tumor cell apoptosis was assessed using a TUNEL assay and HE staining, revealing that the MGC-803 tumor cells in the LV-GFP-CDX2 group had higher levels of apoptosis compared with the LV-GFP-NC and PBS groups (magnification, x400). CDX2, vaudal type homeobox transcription factor 2; LV-GFP, lentivirus-green fluoresent protein; PBS, phosphate-buffered saline; NC, negative control; TV, tumor volume; RTV, relative TV; TUNEL, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling; HE, hematoxylin and eosin.

Article Snippet: Specific rabbit anti-human polyclonal antibodies to CDX2 (#12306), c-Myc (#5605), Skp2 (#2652), Bax (#5023), Bcl-2 (#2827), cyclin D1 (#2978), survivin (#2808) and GAPDH (#2118) were provided by Cell Signaling Technology, Inc. (Beverly, MA, USA) .

Techniques: Over Expression, Control, Injection, TUNEL Assay, Staining, Saline, Negative Control, End Labeling

Journal: Molecular Medicine Reports

Article Title: Overexpression of caudal type homeobox transcription factor 2 inhibits the growth of the MGC-803 human gastric cancer cell line in vivo

doi: 10.3892/mmr.2015.3413

Figure Lengend Snippet: Overexpression of CDX2 mRNA and protein in the LV-GFP-CDX2 group. (A) Reverse transcription semi-quantitative polymerase chain reaction analysis of CDX2 and GAPDH in the MGC-803 tumor tissues from the LV-GFP-CDX2, LV-GFP-NC and PBS groups. M, 500 bp marker. (B) mRNA expression levels of CDX2 were measured in the three groups, normalized to GAPDH and presented as the mean ± standard error of the mean (n=8 in each group). (C) Western blot analysis of the protein expression levels of CDX2 and GAPDH in the MGC-803 tumor tissues from the three groups. (D) Protein expression levels of CDX2 were measured in the three groups, normalized to GAPDH and presented as the mean ± standard error of the mean (n=8 in each group). Lanes: 1, LV-GFP-CDX2 group; 2, LV-GFP-NC group; 3, PBS group, GAPDH: internal control mRNA and protein. * P<0.05 compared with LV-GFP-NC and PBS group, using analysis of variance and Student-Newman-Keuls analyses. CDX2, caudal type homeobox transcription factor 2; LV-GFP, lentivirus-green fluoresent protein; PBS, phosphate-buffered saline; NC, negative control.

Article Snippet: Specific rabbit anti-human polyclonal antibodies to CDX2 (#12306), c-Myc (#5605), Skp2 (#2652), Bax (#5023), Bcl-2 (#2827), cyclin D1 (#2978), survivin (#2808) and GAPDH (#2118) were provided by Cell Signaling Technology, Inc. (Beverly, MA, USA) .

Techniques: Over Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Marker, Expressing, Western Blot, Control, Saline, Negative Control

Journal: Molecular Medicine Reports

Article Title: Overexpression of caudal type homeobox transcription factor 2 inhibits the growth of the MGC-803 human gastric cancer cell line in vivo

doi: 10.3892/mmr.2015.3413

Figure Lengend Snippet: Overexpression of CDX2 induces downregulation in the mRNA expression levels of c-Myc, Skp2, Bcl-2, cyclin D1 and survivin and upregulation in the expression of Bax. (A) Reverse transcription semi-quantitative polymerase chain reaction analysis of c-Myc, Skp2, Bcl-2, cyclin D1, survivin, Bax and GAPDH in the MGC-803 tumor tissues from the LV-GFP-CDX2, 2, LV-GFP-NC and PBS groups. M, 500 bp marker (B) mRNA expression levels of c-Myc, Skp2, Bcl-2, cyclinD1, survivin and Bax were measured in the three groups, normalized to those of GAPDH and presented as the mean ± standard error of the mean (n=8 in each group). 1, LV-GFP-CDX2 group; 2, LV-GFP-NC group; 3, PBS group; GAPDH: internal control. * P<0.05, compared with the LV-GFP-NC and PBS groups, using analysis of variance and Student-Newman-Keuls analyses. Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-associated X protein; Skp2, S-phase kinase-associated protein 2; CDX2, caudal type homeobox transcription factor 2; LV-GFP, lentivirus-green fluoresent protein; PBS, phosphate-buffered saline.

Article Snippet: Specific rabbit anti-human polyclonal antibodies to CDX2 (#12306), c-Myc (#5605), Skp2 (#2652), Bax (#5023), Bcl-2 (#2827), cyclin D1 (#2978), survivin (#2808) and GAPDH (#2118) were provided by Cell Signaling Technology, Inc. (Beverly, MA, USA) .

Techniques: Over Expression, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Marker, Control, Saline

Journal: Molecular Medicine Reports

Article Title: Overexpression of caudal type homeobox transcription factor 2 inhibits the growth of the MGC-803 human gastric cancer cell line in vivo

doi: 10.3892/mmr.2015.3413

Figure Lengend Snippet: Overexpression of CDX2 induces downregulation of the protein expression levels of c-Myc, Skp2, Bcl-2, cyclinD1 and survivin and upregulation of Bax. (A) Western blot analysis of c-Myc, Skp2, Bcl-2, cyclin D1, survivin, Bax and GAPDH in the MGC-803 tumor tissue from the LV-GFP-CDX2, LV-GFP-NC and PBS groups. (B) Protein expression levels of c-Myc, Skp2, Bcl-2, cyclin D1, survivin and Bax were measured in the three groups, normalized to those of GAPDH and expressed as the mean ± standard error of the mean (n=8 in each group). 1, LV-GFP-CDX2 group; 2, LV-GFP-NC group; 3, PBS group; GAPDH: internal control. * P<0.05, compared with the LV-GFP-NC and PBS group, using analysis of variance and Student-Newman-Keuls analyses. Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-associated X protein; Skp2, S-phase kinase-associated protein 2; CDX2, caudal type homeobox transcription factor 2; LV-GFP, lentivirus-green fluoresenct protein; PBS, phosphate-buffered saline.

Article Snippet: Specific rabbit anti-human polyclonal antibodies to CDX2 (#12306), c-Myc (#5605), Skp2 (#2652), Bax (#5023), Bcl-2 (#2827), cyclin D1 (#2978), survivin (#2808) and GAPDH (#2118) were provided by Cell Signaling Technology, Inc. (Beverly, MA, USA) .

Techniques: Over Expression, Expressing, Western Blot, Control, Saline

Journal: Life Science Alliance

Article Title: Laminin is the ECM niche for trophoblast stem cells

doi: 10.26508/lsa.201900515

Figure Lengend Snippet: (A) Expression of undifferentiated TSC marker CDX2 in trophectoderm cells in Lamc1 EQ/+ and Lamc1 EQ/EQ blastocysts. Bar, 50 μm. (B) Loss of integrin α7x2β1 binding to the basement membrane in E5.5 Lamc1 EQ/EQ embryos. Magenta, anti-laminin α1 antibody binding; green, recombinant integrin α7x2β1 binding; white, area double-positive for magenta and green signals. Bar, 50 μm. (C) Morphologies of E5.5 control and Lamc1 EQ/EQ embryos visualized by immunofluorescence. Cyan, laminin α1; magenta, CDX2; green, OCT4. ExE cells are enclosed by dotted lines. The arrow indicates CDX2-positive cells detached from the laminin-positive basement membrane. Bar, 50 μm. (D) Quantification of CDX2-positive and OCT4-positive cells in control WT and Lamc1 EQ/+ and Lamc1 EQ/EQ E5.5 egg cylinders. Data represent means ± SD ( n = 24 and 6 for WT or Lamc1 EQ/+ and Lamc1 EQ/EQ , respectively). *** P < 0.001, significant difference by Welch’s t test. (E) Diagram illustrating the dependence of TSCs on laminin–integrin interactions in the mouse conceptus. In addition to FGF4 and nodal from the epiblast, laminin also acts on TSCs as an ECM niche through binding to integrin receptors. The inset shows the region illustrated in the main figure. The diagram is based on that in .

Article Snippet: The following antibodies and reagents were obtained commercially: rat anti-laminin-γ1 mAb (Millipore); rabbit anti-laminin pAb, BSA, and heparin (Sigma-Aldrich); mouse anti-CDX2 mAb (Biocare); rabbit anti-OCT4 pAb (Santa Cruz Biotechnology); rat anti-integrin αV mAb (RMV-7), hamster anti-integrin β1 mAb (Ha2/5), and hamster anti-integrin β3 mAb (2C9.G2) (BD Biosciences); Alexa 488–conjugated goat antirabbit IgG, Alexa 546–conjugated goat antirat IgG, Alexa 405–conjugated goat antirabbit IgG, Alexa 488–conjugated goat antirat IgG, and Alexa 546–conjugated goat antimouse IgG (Invitrogen); bovine types I and IV collagens (Nippi Inc.); recombinant human FGF4 (Peprotech); and PermaFluor (Thermo Shandon).

Techniques: Expressing, Marker, Binding Assay, Membrane, Recombinant, Control, Immunofluorescence

Journal: Life Science Alliance

Article Title: Laminin is the ECM niche for trophoblast stem cells

doi: 10.26508/lsa.201900515

Figure Lengend Snippet: Morphologies of E5.5 control and Lamc1 EQ/EQ embryos. Cyan, laminin α1; magenta, CDX2; green, OCT4. ExE cells are enclosed by dotted lines. Bar, 50 μm.

Article Snippet: The following antibodies and reagents were obtained commercially: rat anti-laminin-γ1 mAb (Millipore); rabbit anti-laminin pAb, BSA, and heparin (Sigma-Aldrich); mouse anti-CDX2 mAb (Biocare); rabbit anti-OCT4 pAb (Santa Cruz Biotechnology); rat anti-integrin αV mAb (RMV-7), hamster anti-integrin β1 mAb (Ha2/5), and hamster anti-integrin β3 mAb (2C9.G2) (BD Biosciences); Alexa 488–conjugated goat antirabbit IgG, Alexa 546–conjugated goat antirat IgG, Alexa 405–conjugated goat antirabbit IgG, Alexa 488–conjugated goat antirat IgG, and Alexa 546–conjugated goat antimouse IgG (Invitrogen); bovine types I and IV collagens (Nippi Inc.); recombinant human FGF4 (Peprotech); and PermaFluor (Thermo Shandon).

Techniques: Control

Journal: Developmental Cell

Article Title: Maternal DNA Methylation Regulates Early Trophoblast Development

doi: 10.1016/j.devcel.2015.12.027

Figure Lengend Snippet: Scml2 Is Controlled by DNA Methylation and Affects SynT Formation and Cell Adhesion (A) RT-qPCR of Dnmt3a mKO EPCs confirms that Scml2 is controlled by oocyte methylation. (B) Methylation analysis by Sequenom MassARRAY in E7.5 male EPCs, confirming the DMR at an intragenic TSS of Scml2 . Each data point may include more than one CpG from the amplicon, as indicated on the x axis. (C) RT-qPCR analysis of TSCs grown in FGF+ (TSC conditions) or FGF− (differentiation conditions) medium for 6 days, with or without Scml2 overexpression. (D) Expression of Syna is reduced in Dnmt3a mKO EPCs, whereas markers of SynT-II Synb and Cebpa are unaffected. (E) E-cadherin staining of two independent Scml2 knockout clones from TKO TSCs shows a rescue of the morphological alterations seen in TKO TSCs. (F) Scml2 KO on TKO TSCs also rescues the defect in cell adhesion to cell culture wells in the absence of laminin. (G) RT-qPCR analysis of TKO Scml2 KO clones shows maintained expression of the TSC marker Cdx2 ; the expression of genes involved in cell adhesion is not rescued upon Scml2 deletion. Error bars represent SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001; t test comparing WT and Dnmt3a mKO EPCs (A and D) or Scml2 -expressing TSCs versus vector control (C), or ANOVA with post hoc tests comparing Ctrl with DHet/DKO (B) or TKO TSC lines with WT TSCs. See also Figure S6 .

Article Snippet: Paraffin sections were deparaffinized with Histo-Clear and dehydrated through an ethanol series, followed either by standard H&E staining or antigen retrieval by boiling slides for 30 min (in 1 mM EDTA, 0.05% Tween 20, pH 8) and cooling at room temperature for 20 min. After blocking with 1% BSA overnight, sections were incubated with a rabbit monoclonal anti-CDX2 antibody (EPR2764Y, Novus Biologicals, 1:250 dilution) for 2 hr.

Techniques: DNA Methylation Assay, Quantitative RT-PCR, Methylation, Amplification, Over Expression, Expressing, Staining, Knock-Out, Clone Assay, Cell Culture, Marker, Plasmid Preparation, Control

Journal: bioRxiv

Article Title: Calcium signalling is required for anterior patterning in the mouse embryo

doi: 10.1101/2024.11.14.623160

Figure Lengend Snippet: Hex-GFP embryos cultured for 8 hours in the SERCA inhibitor thapsigargin or DMSO control and stained for; (A) epiblast marker OCT4 (DMSO N=4, 10nM thapsigargin N=6) (B) extra-embryonic ectoderm markers CDX2 (DMSO N=5, 10nM thapsigargin N=4), no difference was seen in either marker. (C) Example embryo showing phalloidin staining on cultured embryos showing that the F-actin cytoskeleton is unaffected in thapsigargin cultured embryos (N=18) compared to DMSO (N=15) and that VE cells remain as a monolayer epithelium. Max-intensity projection (MIP).

Article Snippet: Primary antibodies used were: rabbit anti-Cleaved Caspase-3 (Cell Signalling, 9961S, Lot: 43 (06/2014)), rabbit anti-Phospho-Histone H3 (S28) (Cell Signalling, 9713P, Lot: 2 (09/2013)), rabbit anti-OCT-4 (Abcam, ab200834, Lot: GR212680-16), rabbit anti-CDX2 (Cell Signalling, 9775, Lot: 3(12/2017)).

Techniques: Cell Culture, Control, Staining, Marker