zf0808 gd160 cd44 poly cd44 5 r d systems  (R&D Systems)

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