Cells were treated under the indicated conditions, and whole-cell lysates were obtained using radioimmunoprecipitation buffer. The lysates were then subjected to Western blot analysis as reported previously6 (link). The primary antibodies used were anti-cyclin D1, anti-cyclin-dependent kinase 4 (CDK4), anti-glucose-regulated protein 78 (GRP78), anti-ubiquitinated protein, and anti-histone deacetylases (HDACs) 1, 3, and 6 (Santa Cruz Biotechnology); anti-ER-resident protein 44 (ERP44), anti-endoplasmic oxidoreductin-1-like protein (Ero1L), anti-S6 ribosomal protein, and anti-phosphorylated S6 ribosomal protein (Cell Signaling Technology, Danvers, MA, USA); anti-AMP-activated protein kinase (AMPK; Proteintech, Rosemont, IL, USA); anti-acetylated histone (Abcam, Cambridge, UK); and anti-actin (Millipore, Billerica, MA, USA).
Protocol detail
Quantitative Analysis of Protein Expression
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anti-cyclin D1
anti-S6 ribosomal protein
anti-phosphorylated S6 ribosomal protein
anti-acetylated histone
anti-actin
Actin
Amp activated protein kinase
Antibodies
Buffer
Cell
Cyclin d1
Cyclin dependent kinase 4
Ero1l
Glucose
Histone
Histone deacetylases 1
Protein
Radioimmunoprecipitation
Ribosomal protein
Western blot analysis
Corresponding Organization :
Other organizations : National Defense Medical College
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Variable analysis
independent variables
- Cells were treated under the indicated conditions
- Whole-cell lysates were obtained
- Western blot analysis was performed on the lysates
- Radioimmunoprecipitation buffer was used to obtain whole-cell lysates
- No positive or negative controls were explicitly mentioned in the input text.
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