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2 protocols using glut3

1

Phospho-β-catenin Signaling Pathway Modulation

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Canagliflozin, empagliflozin, and dapagliflozin were purchased from Selleck Chemicals (Houston, TX). MG-132, lithium chloride (LiCl), FTY720, WZB117, and phloretin were obtained from Sigma-Aldrich (St. Louis, MO) and Okadaic acid (OA) was from Cayman Chemical (Ann Arbor, MI). For immunoblotting, antibodies against phospho-β-catenin (Ser33/Ser37/Thr41), phospho-β-catenin (Ser45), β-catenin, Cyclin D1, Tubulin, Lamin B, and DDK-tag were purchased from Cell Signaling (Danvers, MA), and Glut1 and GAPDH antibodies were from Abcam (Cambridge, MA). PP2Ac, Glut3, and Actin antibodies were obtained from Millipore (Billerica, MA), Santa Cruz Biotechnology (San Diego, CA), and ProteinTech (Rosemont, IL), respectively. For experiments related to flow cytometry, Monoclonal CD133/2 (293C3)-APC and Mouse-IgG2b-APC antibodies were purchased from Miltenyi Biotech (Bergish Gladbach, Germany), and FITC anti-human CD326 (EpCAM) and Mouse-IgG2b-FITC antibodies were from BioLegend (San Diego, CA).
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2

Immunoblotting Protein Expression Analysis

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Immunoblots were conducted with the following primary antibodies all used at 1:1000: Hexokinase II (cat. no. 2867; Cell Signaling Technology, Danvers, MA, USA), Beta Actin (cat. no. 4970; Cell Signaling), Beta Tubulin (cat. no. 2128; Cell Signaling), GSK3B (cat. no. 9315), p-GSK3B S9 (cat. no. 9323; Cell Signaling), p-p90RSK T573 (cat. no. 9346; Cell Signaling), RSK1/RSK2/RSK3 (cat. no. 9355; Cell Signaling), hsp60 (cat. no. sc-1052; Santa Cruz Biotechnology, Santa Cruz, CA, USA), GLUT1 (cat. no. 07-1401; Millipore, Billerica, MA, USA), GLUT3 (cat. no. NBP1-89762; Novus Biologicals, Littleton, CO, USA) and the secondary antibody Anti-rabbit IgG, HRP-linked Antibody (cat. no. 7074S; Cell Signaling).
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