Journal: Experimental Animals

Article Title: Isolation and morphological characterization of ovine amniotic fluid mesenchymal stem cells

doi: 10.1538/expanim.15-0031

Figure Lengend Snippet: Determination of specific MSC surface markers in oAF-MSCs. Specific markers were detected by immunocytochemistry (ICC). Antibodies against CD13, CD29, CD44, CD90, CD106, and OCT4 showed positive staining, which was indicated by green fluorescence (FITC). CD45 was negative. NC, negative control. Nuclei were stained with DAPI (blue fluorescence).

Article Snippet: The primary antibodies included anti-mouse CD29, CD13, CD44, CD45, CD90, CD106, and OCT4 (Boster Biological Technology, Wuhan, China), and all were diluted with PBS at 1:100.

Techniques: Immunocytochemistry, Staining, Fluorescence, Negative Control

Journal: Cancers

Article Title: Overexpression of Cystine/Glutamate Antiporter xCT Correlates with Nutrient Flexibility and ZEB1 Expression in Highly Clonogenic Glioblastoma Stem-like Cells (GSCs)

doi: 10.3390/cancers13236001

Figure Lengend Snippet: High Gln/Glu ratios predict ZEB1 and xCT expression in primary GBM tumors. ( a ) Expanded regions from 1 H-NMR spectra showing Gln and Glu multiplets of metabolic extracts from seven primary glioblastoma (pGBM) patient samples. Spectra were normalized to the Glu content to visualize the differences in Gln/Glu ratios. ( b ) Immunoblotting of ZEB1, xCT, CD133, CD44, and cMYC protein in GBM patient samples arranged according to their Gln/Glu ratios in ascending order (loading control = β actin). A non-neoplastic brain sample of a trauma patient was blotted as a control. Abbreviations: ctrl, control; Gln, glutamine; Glu, glutamate; n.d., not detectable; pGBM, primary glioblastoma; ppm, parts per million. correspond to b.

Article Snippet: We applied the following primary antibodies and used the indicated dilutions: ZEB1 (1:2000, Sigma, #HPA027524), CD133 (1:250, Miltenyi Biotec, Bergisch Gladbach, Germany, #W6B3C1), c-Myc (1:1000, Thermo Fisher Scientific, #9E10), SOX2 (1:1000, Cell Signaling Technology, Cambridge, UK, #L1D6A2), xCT (1:1000, Thermo Fisher Scientific, #PA1-16893 and 1:1000, Cell Signaling Technologies, #12691), CD44 (1:2000, R&D Systems, Minneapolis, MN, USA, #AF3660), and β-actin (Thermo Fisher Scientific, #MA5-15739 and Cell Signaling Technology, #4970) were incubated overnight at 4 °C in 5% milk powder in Tris-buffered saline + 0.1% Tween-20 (TBST).

Techniques: Expressing, Western Blot, Control

Journal: Journal of Cancer

Article Title: TGF-β2: A Novel Target of CD44-Promoted Breast Cancer Invasion

doi: 10.7150/jca.6638

Figure Lengend Snippet: Time-course western-blot validation of the tetracycline (tet) regulated CD44 expression in MCF7F-B5 breast cancer cell line. Protein lysates were collected at different time points (18, 24 and 48hrs) following withdrawal of tet, in the presence of hyaluronan (HA). The level of expression of CD44s showed a time-dependent increase and was maintained up to 48 hrs.

Article Snippet: The membrane was probed with mouse anti-human CD44s monoclonal antibody (1:500 dilution; R&D Systems,) and then treated with a sheep anti-mouse IgG/horseradish peroxidase conjugate (1:2000 dilution; Santa Cruz Biotechnology).

Techniques: Western Blot, Biomarker Discovery, Expressing