Western blotting was performed as described (8 (link)). Antibodies used were mouse anti-GILZ (G-5) (Santa Cruz Biotechnology), rabbit anti-MKP-1 (Millipore, Bedford, MA), mouse anti-β-actin (Sigma-Aldrich), anti-FoxO3a, anti-TATA-binding protein (TBP), anti-phosphorylated (p) p38, anti-pERK1/2 and total ERK, anti-pAkt and total Akt (Cell Signaling Technology, Danvers, MA) as previously described (8 (link), 30 (link)). Membrane blot density was measured by scanning using the Odyssey system (Li-Cor Biotechnology, Lincoln, NE). Densitometry ratios were normalized to appropriate total protein content and results expressed as a ratio.
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Western Blotting Analysis of Protein Signaling
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Corresponding Organization :
Other organizations : Monash University, University Hospital Cologne, Yale University
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Variable analysis
independent variables
- None explicitly mentioned
- Protein levels/expression of GILZ, MKP-1, β-actin, FoxO3a, TATA-binding protein (TBP), phosphorylated (p) p38, pERK1/2 and total ERK, pAkt and total Akt
- Western blotting procedure as described in previous work (8 and 30)
- Antibodies used: mouse anti-GILZ (G-5) (Santa Cruz Biotechnology), rabbit anti-MKP-1 (Millipore, Bedford, MA), mouse anti-β-actin (Sigma-Aldrich), anti-FoxO3a, anti-TATA-binding protein (TBP), anti-phosphorylated (p) p38, anti-pERK1/2 and total ERK, anti-pAkt and total Akt (Cell Signaling Technology, Danvers, MA)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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