Journal: Diagnostic Pathology

Article Title: Colon adenoma and adenocarcinoma with clear cell components - two case reports

doi: 10.1186/s13000-019-0819-z

Figure Lengend Snippet: Tubular adenoma with clear cell change. The striking tubule structures of the clear cells are accompanied by conventional tubular adenoma cells at low magnification with HE staining ( a ). The boundary between the clear cell and conventional components at high magnification with HE staining ( b ). The clear cell component is negative for PAS ( c ) and alcian blue staining ( d ). Both components are positive for CDX2 staining ( e ). The localization of CEA (f) expression is diffusely cytoplasmic for the clear cell component, and luminal cell apical for the conventional one. Ki67 labeling ( g ) is slightly lower in the clear cell component. d-g represent immunohistochemistry. Ultrastructural examination ( h ) of the boundary between the clear cell area (left) and the conventional adenoma (right) at low magnification is shown and multiple cytoplasmic lipid-like vacuoles surround the nuclei in the clear cells ( i )

Article Snippet: CDX2 , AMT28 , Novus Biologicals, Newcastle, UK , 1:50 , pH 9.0.

Techniques: Staining, Expressing, Labeling, Immunohistochemistry

Journal: Diagnostic Pathology

Article Title: Colon adenoma and adenocarcinoma with clear cell components - two case reports

doi: 10.1186/s13000-019-0819-z

Figure Lengend Snippet: Clear cell adenocarcinoma. Low magnification ( a ) and high magnification of the clear cell ( b ) and conventional components ( c ) with HE staining. The boundary ( d ) between the clear cell and conventional components at high magnification with HE staining. The clear cells are negative for PAS ( e ) and alcian blue staining ( f ), whereas both components of the tumor are positive for CDX2 ( g ). The localization of CEA ( h ) expression is diffusely cytoplasmic for the clear cell component and luminal cell apical for the conventional component. CD10 ( i ) and adipophilin ( j ) expression is confined to the clear component and Ki67 labeling ( k ) is higher in the clear cell component. g-k represent immunohistochemistry. Immunoelectron microscopy analysis ( l ) at low (left) and high magnification (right) reveals multiple cytoplasmic lipid-like vacuoles in clear cells that are negative for adipophilin

Article Snippet: CDX2 , AMT28 , Novus Biologicals, Newcastle, UK , 1:50 , pH 9.0.

Techniques: Staining, Expressing, Labeling, Immunohistochemistry, Immuno-Electron Microscopy

Journal: Diagnostic Pathology

Article Title: Colon adenoma and adenocarcinoma with clear cell components - two case reports

doi: 10.1186/s13000-019-0819-z

Figure Lengend Snippet: Antibody information

Article Snippet: CDX2 , AMT28 , Novus Biologicals, Newcastle, UK , 1:50 , pH 9.0.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Direct On-Chip Differentiation of Intestinal Tubules from Induced Pluripotent Stem Cells

doi: 10.3390/ijms21144964

Figure Lengend Snippet: Characterization of hiPSC-derived human intestinal-like tubules. ( A ) 3D reconstruction image of an iPSC-derived tubule at Day 4 stained with antibodies for SOX17 (green), FOXA2 (red) or DAPI for DNA (blue). Cells attach and form tubule at the DE stage. ( B ) Representative images of iPSC tubules stained for Definitive endoderm markers SOX17 and FOXA2 at day 4. ( C ) Hindgut marker CDX2 at day 7. ( D ) Gene expression was measured using TaqMan qRT-PCR from hiPSC tubules at different differentiation stages. The following genes were analysed: POU class 5 homeobox 1 (POU5F1, indicating pluripotency); Nanog homeobox (NANOG, indicating Primitive Streak):, forkhead box a2 (FOXA2, Definitive Endoderm), SRY (sex determining region Y)-box 17 (SOX17, Definitive Endoderm), and Homeobox protein CDX-2 (Posterior gut). ( E ) Representative images of iPSC tubules stained for intestinal markers Lysozyme (LYZ), Villin (VIL) and Chromogranin A (CHGA) at day 28 (green). Nuclei were stained with DAPI (blue) to visualize the overall morphology. Scale bars = 100 µm. ( F ) Relative gene expression analysis of Intestinal markers: Leucine-rich repeat-containing G-protein coupled receptor 5 (LGR5), Mucin-2 (MUC2), Lysozyme (LYZ), Villin-1 (VIL1), Chromagranin A (CHGA) and sucrase-isomaltase (SI). ( G ) Relative mRNA expression of MDR1 (P-gp) and CYP3A4 in iPSC-derived intestine-like tubules on day 14, day 28 and day 31, human adult colon organoids and Caco-2 cells. Expression levels were normalized to beta-actin (ACTB), data represented as mean ± SD relative to the expression in undifferentiated miFF1 hiPSC ( n = 2, n ≥ 2–3). The Y-axis represents the LOG10 relative quantification (RQ). Caco-2 cells and primary colon organoid were also included to compare gene expression and to follow the differentiation of our model during the different stages. Data are presented as the average of two independent experiments +/− SD ( n = 3).

Article Snippet: 1/100) CDX2 (R&D Systems, AF3665, 1/100), Villin (Santa Cruz, 58897, 1/200), Chromagranin A (Rabbit Santa, cruz13090, 1/200), Lysozyme (Thermo Fischer, MA1-82873,1/200), Isotype Rabbit (Life Technologies, 86199, 1/250) Isotype Goat (Life Technologies, 026202, 1/250), Isotype Mouse (Invitrogen, 08-6599 1/250), Goat Alexa Fluor 488 (Invitrogen, A11055 1/250) Mouse Alexa Fluor 555(Life Technologies, 1736967, 1/250), Rabbit Alexa Fluor 647 (MERCK, SAB460177 1/250) and cell stains Actin Green (LifeTechnologies, R37110), Hoechst 33342 (Thermo Fischer, H3570, 1/1000).

Techniques: Derivative Assay, Staining, Marker, Gene Expression, Quantitative RT-PCR, Expressing, Quantitative Proteomics

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