Quantitative Immunoblot Analysis of Cell Signaling

Quantitative immunoblot analyses were conducted using standard procedures [16 (link)]. Antibodies directed to ERK1/2 (#9102), phospho-ERK1/2 (#9101), VEGFR2 (#9698), phospho-VEGFR2 Tyr1115 (#3770), acetyl-histone H4 Lys8 (#2594), phospho-AKT Thr308 (#4056), phospho-AKT Ser473 (#3787), AKT (#4691), phospho-FOXO1 Ser256 (#9461), FOXO1 (#2880), phospho-4EBP1 Thr37/46 (#2855), phospho-S6K Ser235/236 (#4858) were purchased from Cell Signaling Technology and used at dilutions of 1:1000. Antibodies to RET (# ab134100) and phospho-RET Tyr 1062 (#ab51103) were purchased from Abcam and used at dilutions of 1:10,000 and 1:2000 respectively. Antibodies to Actin (#A2228) were from Sigma-Aldrich and used at dilution 1:10,000.

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Pozo K., Zahler S., Ishimatsu K., Carter A.M., Telange R., Tan C., Wang S., Pfragner R., Fujimoto J., Grubbs E.G., Takahashi M., Oltmann S.C, & Bibb J.A. (2018). Preclinical characterization of tyrosine kinase inhibitor-based targeted therapies for neuroendocrine thyroid cancer. Oncotarget, 9(102), 37662-37675.

Publication 2018

phospho-ERK1/2 VEGFR2 acetyl-histone H4 Lys8 phospho-AKT Thr308 phospho-AKT Ser473 phospho-FOXO1 Ser256 FOXO1 phospho-4EBP1 Thr37/46 Antibodies to Actin

4ebp1 Actin Antibodies Dilution Erk1 Histone h4 Immunoblot analyses Vegfr2

Corresponding Organization : University of Alabama at Birmingham

Other organizations : The University of Texas Southwestern Medical Center, Ludwig-Maximilians-Universität München, Advanced Imaging Research (United States), Medical University of Graz, The University of Texas MD Anderson Cancer Center

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Levels of ERK1/2
Levels of phospho-ERK1/2
Levels of VEGFR2
Levels of phospho-VEGFR2 Tyr1115
Levels of acetyl-histone H4 Lys8
Levels of phospho-AKT Thr308
Levels of phospho-AKT Ser473
Levels of AKT
Levels of phospho-FOXO1 Ser256
Levels of FOXO1
Levels of phospho-4EBP1 Thr37/46
Levels of phospho-S6K Ser235/236
Levels of RET
Levels of phospho-RET Tyr 1062
Levels of Actin

control variables

None explicitly mentioned

positive controls

None explicitly mentioned

negative controls

None explicitly mentioned

Annotations

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