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3 protocols using anti aldh1a1

1

Investigating Sorafenib and Cellular Signaling

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Sorafenib was purchased from Sigma‐Aldrich (St. Louis, MO, USA). The compound A‐769662 and capsaicin were purchased from Tocris Bioscience (Bristol, UK). The primary antibodies anti‐CD133, anti‐ALDH1A1, anti‐pAkt‐ser473, p‐mTOR, pAMPKα1‐thr172, pACC‐ser79, anti‐cyclin D1, anti‐PGC1α, and anti‐PPARγ and the antibodies against the corresponding total forms were obtained from Cell Signaling Technology (Danvers, MA, USA). The primary antibody anti‐β‐catenin was purchased from Santa Cruz Biotechnology (Dallas, Texas, USA). The primary antibody anti‐Hif‐1α was purchased from Novus (St. Louis, MO, USA). Anti‐alpha fetoprotein and peroxidase‐labelled secondary anti‐mouse IgG were purchased from Sigma‐Aldrich, and anti‐rabbit IgG was purchased from Calbiochem (San Diego, USA).
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2

Western Blot Analysis of Cellular Proteins

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Whole-cell lysates were prepared by boiling cell pellets for 10 min in SDS lysis buffer [2% SDS, 10% glycerol, 62 mmol/L Tris-HCl, pH 6.8, and a complete mini-protease inhibitor mixture (Roche Applied Science)]. After protein quantification, equal amounts of proteins were loaded, separated on a polyacrylamide gel, and transferred to a nitrocellulose membrane. Protein bands were immuno-detected with appropriate antibodies: anti-ALDH1A1 (Cell Signaling, #54135), anti-BRD4 (Cell Signaling, #13440), anti-β-Tubulin (Cell Signaling, #2148), and anti-GAPDH (Santa Cruz, Sc-47724).
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3

Scoulerine Inhibits Ovarian Cancer

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Scoulerine was procured from Herbest Bio-Tech, China (CAS 6451-73-6) (purity > 98 %). Human ovarian epithelial adenocarcinoma cells (OVCAR3; ATCC ® HTB161™) and HEK293; ATCC ® CRL-1573™) were obtained from the ATCC, USA. Antibodies, anti-GAPDH, anti-N-Cadherin, anti-E-Cadherin, anti-Vimentin, anti-CD44, anti-Oct-4, anti-Sox2, anti-Ald-h1A1, anti-Akt and anti-p-Akt were obtained from Cell Signaling Technology (USA). While other antibodies, anti-CD24, anti-PI3K, anti-mTOR and anti-p-mTOR were purchased from Santa Cruz Biotechnology (USA). Clarity Western ECL substrate luminol assay kit was obtained from (BioRad Laboratories, Inc. Tumorsphere medium was obtained from Stem Cell Technologies. All chemicals are purchased from Sigma or stated accordingly.
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