Immunoblotting and Nuclear Fractionation
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Corresponding Organization : University of Chicago
Other organizations : Keio University, Duke University, University of Wisconsin–Madison
Variable analysis
- Cell lysates were prepared in RIPA buffer
- Nuclear fractionation was performed using the REAP cellular fractionation method
- Immunoblotting was performed as previously described (19)
- Levels of SPHK1, SGPL1, HIF1α, HIF2α, phospho-AKT (S473), AKT, SOX2, phospho-AMPK (T172), AMPK, c-MYC, OCT-1, PKM1/2, cleaved caspase 3, Snail, and V5-tag were measured
- β-Tubulin, Histone H3, and β-Actin were used as loading controls
- Positive control: Not specified
- Negative control: Not specified
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