Signaling Pathway Analysis Protocol

Western blot, cell fractionation and Co-IP analysis were carried out as described in our previous publications.^{23 (link),26 (link),29 (link)} The following primary antibodies were used: anti-ITGA9, anti-CD31 (Abcam, Cambridge, MA); anti-β-catenin, anti-non-phospho-β-catenin, phospho-β-catenin (Ser33/37/Thr41, Thr41/Ser45, Ser675), phospho-GSK3α (Ser21), phospho-GSK3β (Ser9), total GSK3α, total GSK3β, phosphor-CREB (Ser133), total CREB, integrin-linked kinase (ILK), PKA-Cα, and PARP-1 (Cell Signaling Technology, Beverly, MA); anti-PKA RIIα regulatory unit (Santa Cruz Biotechnology, Dallas, TX); anti-β-actin (Sigma, St. Louis, MO).

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Wang Z., Li Y., Xiao Y., Lin H.P., Yang P., Humphries B., Gao T, & Yang C. (2019). Integrin α9 depletion promotes β-catenin degradation to suppress triple negative breast cancer tumor growth and metastasis. International journal of cancer, 145(10), 2767-2780.

Publication 2019

anti-CD31 anti-β-catenin anti-non-phospho-β-catenin phospho-β-catenin phospho-GSK3α phospho-GSK3β total GSK3α total GSK3β phosphor-CREB total CREB PKA-Cα PARP-1 anti-β-actin

Actin Antibodies Cell fractionation Gsk3α Gsk3β Integrin linked kinase Itga9 Parp 1 Phosphor Western blot β catenin

Corresponding Organization : University of Kentucky

Other organizations : TCM-Intigrated Cancer Center of Southern Medical University, Guangzhou Medical University, University of Michigan–Ann Arbor

Top 5 similar protocols

Variable analysis

independent variables

Western blot
Cell fractionation
Co-IP analysis

dependent variables

ITGA9 expression
CD31 expression
β-catenin expression and phosphorylation
GSK3α and GSK3β expression and phosphorylation
CREB expression and phosphorylation
ILK expression
PKA-Cα expression
PARP-1 expression

control variables

Experimental procedures described in previous publications

controls

Positive controls: Not explicitly mentioned.
Negative controls: Not explicitly mentioned.

Annotations

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