Western Blot Analysis of Cell Signaling

Following gene transfection or drug treatments, cell lysates were prepared and quantified before performing further experiments. Western blotting analysis was carried out as previously described (Wang et al, 2013a (link)). The following antibodies were used: anti-ROC1 (Abcam, Cambridge, MA, USA); anti-Phospho-4E-BP1 (Thr37/46) and anti-E-cadherin (Cell Signaling); anti-non-4E-BP1 and anti-glyceraldehyde-3-phosphate dehydrogenase (anti-GAPDH; Epitomics, Hangzhou, China); anti-vimentin and anti-N-cadherin (Santa Cruz); and anti-DEPTOR (Sigma).

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Wang W., Chen H., Liu Z., Qu P., Lan J., Chen H., Zou L, & Qiu J. (2016). Regulator of cullins-1 expression knockdown suppresses the malignant progression of muscle-invasive transitional cell carcinoma by regulating mTOR/DEPTOR pathway. British Journal of Cancer, 114(3), 305-313.

Publication 2016

anti-ROC1 anti-E-cadherin

4e bp1 Antibodies Cadherin Cell Drug Gapdh Gene Glyceraldehyde 3 phosphate dehydrogenase Transfection Vimentin Western blotting analysis

Corresponding Organization :

Other organizations : Nantong University, Affiliated Hospital of Nantong University, Yancheng First People's Hospital, Shanghai First People's Hospital, Shanghai Jiao Tong University

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Protocol cited in 1 other protocol

Variable analysis

independent variables

Gene transfection
Drug treatments

dependent variables

Protein expression of ROC1
Phosphorylation of 4E-BP1 (Thr37/46)
Expression of E-cadherin
Expression of vimentin
Expression of N-cadherin
Expression of DEPTOR

control variables

Cell lysates were prepared and quantified before performing further experiments
Western blotting analysis was carried out as previously described (Wang et al, 2013a)

positive controls

Anti-non-4E-BP1
Anti-glyceraldehyde-3-phosphate dehydrogenase (anti-GAPDH)

negative controls

None specified

Annotations

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