HSP90 inhibitors 17-AAG, luminespib, and ganetespib were obtained from SelleckChem (Houston, TX). CHX and MG132 were purchased from Sigma-Aldrich (St. Louis, MO), and rhIL-8 was purchased from PeproTech (Rocky Hill, NJ). MTS Assay Kit was obtained from Abcam (Cambridge, UK). Glycolysis antibody sampler kit, including PKM2, PFKP, glyceraldehyde-3-phosphate dehydrogenase, hexokinase I (HKI), HKII, lactate dehydrogenase A (LDHA), and PDH antibodies, was purchased from Cell Signaling Technology (Beverly, MA). Antibodies that recognize Ub, PDL1, and IL-8 were purchased from Novus Biologicals (Littleton, CO). Antibodies against IL-1α and IL-1β were ordered from Abcam, and antibodies against HSP90 and β-actin were obtained from Sigma-Aldrich (St. Louis, MO). Antibody that recognizes CD8α was purchased from Cell Signaling Technology. Secondary antibodies were purchased from Invitrogen (Carlsbad, CA). Cell proliferation, viability, colony formation, and Western blot assays were carried out as previously described (18 (link), 43 (link)).
Protocol detail
Investigating HSP90 Inhibitor-Mediated Glycolytic Changes
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Corresponding Organization : Georgia Institute of Technology
Other organizations : University of Georgia, Columbia University, Columbia University Irving Medical Center
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Variable analysis
independent variables
- 17-AAG
- Luminespib
- Ganetespib
- MG132
- RhIL-8
- Cell proliferation
- Cell viability
- Colony formation
- Protein expression levels (PKM2, PFKP, glyceraldehyde-3-phosphate dehydrogenase, HKI, HKII, LDHA, PDH, Ub, PDL1, IL-8, IL-1α, IL-1β, HSP90, CD8α, β-actin)
- MTS Assay Kit
- Glycolysis antibody sampler kit
- Antibodies against Ub, PDL1, IL-8, IL-1α, IL-1β, HSP90, β-actin, CD8α
- Secondary antibodies
- Positive controls: Not explicitly mentioned.
- Negative controls: Not explicitly mentioned.
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