Real-time PCR analysis was performed with probe sets Stk11 (Mm00488473_g1), Hdc (Mm00456104_m1), Ctnnb1 (Mm00483039_m1), and Actb (Mm00607939_s1) (all from ThermoFisher Scientific). Immunoblots were performed and quantified as described previously34 (link), using the following antibodies: phospho-LKB1 (Ser428; C67A3), LKB1 (D60C5), β-catenin (D10A8), phospho-β-catenin (Ser33/37/Thr41), phospho-S6 (Ser235/236; D57.2.2E), phospho-AKT (Ser473; D9E), phospho-FOXO1 (Ser256), phospho-AMPK (Thr172) (all from Cell Signaling Technology), and β-ACTIN (Sigma).
Protocol detail
Quantitative Analysis of Cellular Signaling Pathways
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Corresponding Organization :
Other organizations : St. Jude Children's Research Hospital, Broad Institute, Harvard University, Brigham and Women's Hospital
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Variable analysis
independent variables
- Stk11
- Ctnnb1
- Phospho-LKB1 (Ser428)
- β-catenin
- Phospho-β-catenin (Ser33/37/Thr41)
- Phospho-S6 (Ser235/236)
- Phospho-AKT (Ser473)
- Phospho-FOXO1 (Ser256)
- Phospho-AMPK (Thr172)
- β-ACTIN
- Positive control: None specified.
- Negative control: None specified.
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