Western Blotting Analysis of EMT Markers

Standard western blot analysis was carried out as described previously [26 (link)]. Antibodies against E-cadherin (ab76055), vimentin (ab92547), VEGF (vascular endothelial growth factor, ab46154) and TGF-β1(ab25121) were purchased from Abcam (Cambridge, MA). The following antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA): Smad2/3 (#3102), p-Smad2 (#3108), p-Smad3 (#9520), MMP-2 (matrix metalloproteinase-2, #4022) and MMP-9 (#3852). SHCBP1 antibody (12672–1-AP) was purchased from Proteintech (Wuhan, China). β-actin (sc-8432) antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

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Peng C., Zhao H., Song Y., Chen W., Wang X., Liu X., Zhang C., Zhao J., Li J., Cheng G., Wu D., Gao C, & Wang X. (2017). SHCBP1 promotes synovial sarcoma cell metastasis via targeting TGF-β1/Smad signaling pathway and is associated with poor prognosis. Journal of Experimental & Clinical Cancer Research : CR, 36, 141.

Publication 2017

ab76055 ab92547 ab46154 ab25121 Smad2/3 p-Smad2 p-Smad3 MMP-2 MMP-9

Actin Antibodies Antibody Cadherin Matrix metalloproteinase 2 Mmp 2 Mmp 9 Smad2 Smad3 Tgf β1 Vascular endothelial growth factor Vimentin Western blot analysis

Corresponding Organization :

Other organizations : Second Hospital of Shandong University, Capital Medical University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Expression levels of E-cadherin, vimentin, VEGF, TGF-β1, Smad2/3, p-Smad2, p-Smad3, MMP-2, and MMP-9

control variables

None explicitly mentioned

controls

Positive control: None explicitly mentioned
Negative control: None explicitly mentioned

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