Journal: Biology of reproduction

Article Title: Expression of two isoforms of CD44 in human endometrium.

doi: 10.1095/biolreprod51.4.739

Figure Lengend Snippet: FIG. 1. Immunofluorescence of CD44 in human endometrium. a) Proliferative phase tissue (mAb P3H9) showing immunoreactivity in epithelial cells in a gland as well as in surrounding stroma. b) Decidua of first trimester (mAb P3H9) with strong reactivity in decidual stromal cells. c) Late secretory phase tissue (mAb PIG12) showing lateral staining in gland cells. Gland lumen is to right of the field. d) Nuclear staining in same sections as c. e) Example of tissue (late secretory phase) in which stromal reactivity is present but many glands are unstained (mAb PIG12). Note that one gland at bottom right is immunopositive. f) Nuclear staining, same field as (e). Arrowheads in a, c, and e indicate the apical cell surface of glandular epithelium.

Article Snippet: MATERIALS AND METHODS Antibodies Mouse pan anti-CD44 monoclonal antibodies (mAbs) were as follows: P1G12 and P3H9 (14,15; Chemicon, Temecula, CA); F10-44-2 (Serotec, Oxford, UK); BU52 (The Binding Site, Birmingham, UK); A1G3 (Seralab, High Wycombe, UK); and E1.2, a generous gift from Dr. C. Isacke, Imperial College, London [38].

Techniques: Immunofluorescence, Staining

Journal: Biology of reproduction

Article Title: Expression of two isoforms of CD44 in human endometrium.

doi: 10.1095/biolreprod51.4.739

Figure Lengend Snippet: FIG. 2. Immunofluorescence of CD44 in endometrial epithelial cells (mAb P3H9). al) Freshly isolated gland fragment in confocal microscopy. b) Pri- mary culture at 4 days showing mainly lateral immunoreactivity. c) Ishi- kawa endometrial adenocarcinoma cells with lateral as well as punctate surface staining.

Article Snippet: MATERIALS AND METHODS Antibodies Mouse pan anti-CD44 monoclonal antibodies (mAbs) were as follows: P1G12 and P3H9 (14,15; Chemicon, Temecula, CA); F10-44-2 (Serotec, Oxford, UK); BU52 (The Binding Site, Birmingham, UK); A1G3 (Seralab, High Wycombe, UK); and E1.2, a generous gift from Dr. C. Isacke, Imperial College, London [38].

Techniques: Immunofluorescence, Isolation, Confocal Microscopy, Staining

Journal: Biology of reproduction

Article Title: Expression of two isoforms of CD44 in human endometrium.

doi: 10.1095/biolreprod51.4.739

Figure Lengend Snippet: FIG. 3. Immunoprecipitation of CD44 from surface iodinated gland cells with mAb PIG12 gives a band at approximately 130 kDa. Right lane: con- trol, irrelevant mAb. Dots at right indicate positions of molecular size mark- ers (from the top down) of 200, 116, 67, and 45 kDa.

Article Snippet: MATERIALS AND METHODS Antibodies Mouse pan anti-CD44 monoclonal antibodies (mAbs) were as follows: P1G12 and P3H9 (14,15; Chemicon, Temecula, CA); F10-44-2 (Serotec, Oxford, UK); BU52 (The Binding Site, Birmingham, UK); A1G3 (Seralab, High Wycombe, UK); and E1.2, a generous gift from Dr. C. Isacke, Imperial College, London [38].

Techniques: Immunoprecipitation

Journal: International journal of oncology

Article Title: Epithelial membrane protein 3 regulates lung cancer stem cells via the TGF‑β signaling pathway.

doi: 10.3892/ijo.2021.5261

Figure Lengend Snippet: Figure 2. EMP3 regulates CSC properties in lung CSC. (A) Western blot analysis of the CSC marker proteins CD133, ALDH1A1, ALDH1A3 and CD44. The CSC regulatory proteins Sox2, Oct‑4 and Nanog were also analyzed by western blotting. A549 cells were transfected with siRNA targeting EMP3. (B) Immunocytochemistry analysis of CSC marker proteins using siRNA treated A549 cells. The green signal is produced by the GFP tag on the secondary antibody. (C) Sphere‑forming capacity analysis of A549 cells in siRNA transfected EMP3 cells and (D) the ability to of an anti‑EMP3 antibody to abrogate sphere formation was assessed. (E) Single‑cell assay of si‑EMP3 transfected cells and (F) the ability of the anti‑EMP3 antibody to abrogate this. (G) Limiting dilution assays were performed in 96 well plates. Wells were plated with 1, 50, 100, 150 or 200 cells/well, and conditioned media was added. Results were confirmed 10 days after seeding of cells. Colony‑formation assays to observe the clonogenicity of the (H) EMP3‑knockdown A549 and (I) anti‑EMP3 antibody treated A549 cells. Cells were irradiated with 3 Gy radiation. After 10 days of incubation, the colonies were stained with crystal violet. (J) Western blot analysis of members of the Sonic hedgehog, Wnt/β‑catenin and Notch signaling pathways in CSCs. Data are presented as the mean ± standard deviation of three repeats. Scale bar, 50 µm. *P<0.05 vs. control. EMP3, epithelial membrane protein 3; ALDH1, aldehyde dehydrogenase 1; CSC, cancer stem cell; Sox2, sex determining region Y‑box 2; Oct‑4, octamer‑binding transcription factor 4; siRNA, small interfering RNA; p, phosphorylated; GSK3‑β, glycogen synthase kinase 3‑β.

Article Snippet: Antibodies against EMP3 (cat. no. ab236671; Abcam), Sox2 (sex determining region Y‐box 2; cat. no. 3579; Cell Signaling Technology, Inc.), phos‐ phorylated (p)‐Smad2 (cat. no. 18338; Cell Signaling Technology, Inc.), p‐Smad3 (cat. no. 9520; Cel l Signaling Technology, Inc.), Smad2/3 (cat. no. 5678; Cell Signaling Technology, Inc.), β‐actin (cat. no. sc‐7963; Santa Cruz Biotechnology, Inc.), TGFBR1 (cat. no. sc‐518086; Santa Cruz Biotechnology, Inc.), TGFBR2 (cat. no. sc‐17791; Santa Cruz Biotechnology, Inc.), TGFBR3 (cat. no. sc‐74511; Santa Cruz Biotechnology, Inc.), CD44 (cat. no. 5640; Cell Signaling Technology, Inc.), β‐catenin (cat. no. sc‐7963; Santa Cruz Biotechnology, Inc.), Twist (cat. no. sc‐15393; Santa Cruz Biotechnology, Inc.), Vimentin (MA5‐16409; Thermo Fisher Scientific, Inc.), alde‐ hyde dehydrogenase (ALDH1)A1 (cat. no. ab52492; Abcam), ALDH1A3 (cat. no. ab129815; Abcam), Snail (cat. no. sc‐10432; Santa Cruz Biotechnology, Inc.), Slug (cat. no. sc‐166476; Santa Cruz Biotechnology, Inc.), ZEB1 (Zinc finger E‐box‐binding homeobox 1; cat. no. sc‐25388; Santa Cruz Biotechnology, Inc.), E‐cadherin (cat. no ab15148; Abcam), N‐cadherin (cat. no. 610921; BD Biosciences) and Oct4 (cat. no. 2750; Cell Signaling Technology, Inc.) were used.

Techniques: Western Blot, Marker, Transfection, Immunocytochemistry, Produced, Irradiation, Incubation, Staining, Protein-Protein interactions, Standard Deviation, Control, Membrane, Small Interfering RNA

Journal: STAR Protocols

Article Title: An optimized protocol for phenotyping human granulocytes by mass cytometry

doi: 10.1016/j.xpro.2022.101280

Figure Lengend Snippet:

Article Snippet: CD44 (Clone IM7) - 171Yb , Fluidigm Sciences , Cat#3171003B.

Techniques: Recombinant, Blocking Assay, Electron Microscopy, Red Blood Cell Lysis, Antibody Labeling, Membrane, Software

Journal: STAR Protocols

Article Title: An optimized protocol for phenotyping human granulocytes by mass cytometry

doi: 10.1016/j.xpro.2022.101280

Figure Lengend Snippet: Mass cytometry surface antibody panel

Article Snippet: CD44 (Clone IM7) - 171Yb , Fluidigm Sciences , Cat#3171003B.

Techniques: Mass Cytometry

Journal: Stem Cell Research & Therapy

Article Title: Glioma-associated mesenchymal stem cells-mediated PD-L1 expression is attenuated by Ad5-Ki67/IL-15 in GBM treatment

doi: 10.1186/s13287-022-02968-z

Figure Lengend Snippet: Identification of MSCs derived from human glioma tissues. A and B Adherent growth patterns and fibroblastic morphology of GA-MSCs cultured in MSC media (× 100, scale bars = 1000 µm). C Immunofluorescence showed that GA-MSCs expressed MSC specific markers CD44 and CD105 (× 400, scale bars = 500 µm)

Article Snippet: The cells were incubated with primary antibodies against CD44 and CD105 (1:100, Proteintech, China), then overnight at 4 °C.

Techniques: Derivative Assay, Cell Culture, Immunofluorescence

Journal: International journal of stem cells

Article Title: Collagen Scaffold Augments the Therapeutic Effect of Human Umbilical Cord Mesenchymal Stem Cells in a Rat Model of Intrauterine Adhesion.

doi: 10.15283/ijsc24079

Figure Lengend Snippet: Fig. 1. Characterization of human umbilical cord mesenchymal stem cells (hUCMSCs). (A) Morphology of fourth-generation hUCMSCs under light microscope, red arrows represent the typical parts, the magnification is 10×. (B, C) Detection of multidirectional differentiation potential (B into chondrogenic differentiation, C into adipogenic differentiation) characteristic of hUCMSCs, red arrows represent the typical parts, the magnifications are 10×. (D-I) Flow cytometry detection of positive markers (CD29, CD105, CD90, CD44) and negative markers (CD45, CD34) characteristic of human umbilical cord MSCs.

Article Snippet: The cells were incubated with 5 μL of antihuman CD105, CD34, CD45, CD29, CD44, and CD90 antibodies (Elabscience) in the dark for 30 minutes.

Techniques: Light Microscopy, Flow Cytometry

Journal: PLoS ONE

Article Title: CD166/ALCAM Expression Is Characteristic of Tumorigenicity and Invasive and Migratory Activities of Pancreatic Cancer Cells

doi: 10.1371/journal.pone.0107247

Figure Lengend Snippet: (A) qRT-PCR analysis of the mRNA levels of CD166 in SUIT-2 cells after RNA interference was performed at the indicated days post-transfection. Control (siControl) or CD166 silenced cells (siCD166) were analyzed by (B) colony formation assays at the indicated days post-transfection. (C) qRT-PCR analysis of EMT markers E-cadherin, N-cadherin, Zeb-1, and MMP2 in CD166+ and CD166- Panc-1 and SW1990 cells. (D) The relationships between expression of CD166 and CSC markers CD24, CD44, and CD133 in Panc-1 cells were analyzed by flow cytometry. Data represent the mean ± SD; *, p <0.05; NS, not significant.

Article Snippet: Cells from subconfluent monolayer cultures were suspended in phosphate-buffered saline (PBS) and incubated with monoclonal anti-human ALCAM-phycoerythrin (PE) (R&D Systems, Minneapolis, MN), anti-human CD24-fluorescein isothiocyanate (FITC) (eBioscience Inc, San Diego, CA), anti-human CD44-FITC (MBL, Nagoya, Japan), and monoclonal anti-human CD133-FITC (Ancell Corp, Bayport, MN) antibodies.

Techniques: Quantitative RT-PCR, Transfection, Expressing, Flow Cytometry