Western Blotting Analysis of Cell Signaling Proteins

Western blotting was performed using a SDS-PAGE Electrophoresis System according to the previous^{39 (link), 40 (link)} description with rabbit polyclonal anti-FAP antibody (1 : 1000; LifeSpan BioSciences Inc., Seattle, WA, USA), anti-β-actin, CCND1, CDK4, p21, E2F1, and p27 (1 : 500; Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-C-Myc, PTEN, β-catenin, GSK-3b, p-GSK-3b(ser-9), p-Erk1/2(Tyr202/Tyr204),Erk1/2, p-MEK1/2 (Ser-217/221), MEK1,CCNE1, MMP2, MMP9, p-RB (ser780), RB, AKT, p-Akt (Ser-473), PI3K, p-PI3K (Tyr458), p16, p27, Slug, Snail, Vimentin, N-cadherin, and E-cadherin (1 : 1000; Cell Signaling Technology, Danvers, MA, USA). Signals were detected using enhanced chemiluminescence reagents (Pierce, Rockford, IL, USA).

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Wang H., Wu Q., Liu Z., Luo X., Fan Y., Liu Y., Zhang Y., Hua S., Fu Q., Zhao M., Chen Y., Fang W, & Lv X. (2014). Downregulation of FAP suppresses cell proliferation and metastasis through PTEN/PI3K/AKT and Ras-ERK signaling in oral squamous cell carcinoma. Cell Death & Disease, 5(4), e1155-.

Publication 2014

anti-β-actin CCND1 anti-C-Myc β-catenin GSK-3b p-GSK-3b(ser-9), p-Erk1/2 Erk1/2 p-MEK1/2 MMP2 MMP9 Snail Vimentin N-cadherin E-cadherin

Actin Anti antibody Ccnd1 Ccne1 Chemiluminescence E cadherin E2f1 Electrophoresis Erk1 Mek1 Mmp2 Mmp9 N cadherin Pi3k Pten Rabbit Sds page Slug Snail Vimentin β catenin

Corresponding Organization :

Other organizations : Southern Medical University, Nanfang Hospital

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Variable analysis

independent variables

SDS-PAGE Electrophoresis System

dependent variables

Signals detected using enhanced chemiluminescence reagents

control variables

Rabbit polyclonal anti-FAP antibody (1 : 1000; LifeSpan BioSciences Inc., Seattle, WA, USA)
Anti-β-actin, CCND1, CDK4, p21, E2F1, and p27 (1 : 500; Santa Cruz Biotechnology, Santa Cruz, CA, USA)
Anti-C-Myc, PTEN, β-catenin, GSK-3b, p-GSK-3b(ser-9), p-Erk1/2(Tyr202/Tyr204),Erk1/2, p-MEK1/2 (Ser-217/221), MEK1,CCNE1, MMP2, MMP9, p-RB (ser780), RB, AKT, p-Akt (Ser-473), PI3K, p-PI3K (Tyr458), p16, p27, Slug, Snail, Vimentin, N-cadherin, and E-cadherin (1 : 1000; Cell Signaling Technology, Danvers, MA, USA)

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