Protein Expression Analysis by Western Blot

Cellular protein lysates were prepared by homogenization with 1× RIPA lysis buffer (Cell Signaling Technology, Billerica, MA). Protein concentrations were measured using a protein assay dye (Bio-Rad Laboratories, Hercules, CA). SDS-PAGE and immunoblotting analysis were performed as described previously^{3 (link)}. The detecting antibodies were raised against E-cadherin (ab76055, abcam), vimentin (2707-1 epitomics), α-SMA (ab5694, abcam), fibronectin (ab2413, abcam), WNT5A (MA5-15502, Invitrogen), β-catenin (13-8400, Invitrogen), NF-κB (ab32536, abcam), MMP-9 (13667, CST), and GAPDH (GTX627408, GeneTex).

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Chung Y.H., Cheng Y.T., Kao Y.H., Tsai W.C., Huang G.K., Chen Y.T., Shen Y.C., Tai M.H, & Chiang P.H. (2022). MiR-26a-5p as a useful therapeutic target for upper tract urothelial carcinoma by regulating WNT5A/β-catenin signaling. Scientific Reports, 12, 6955.

Publication 2022

1× RIPA lysis buffer protein assay dye ab76055 ab5694 ab2413 MA5-15502 ab32536 GTX627408

A protein Antibodies Assay Buffer Cadherin Cellular Fibronectin Gapdh Immunoblotting analysis Mmp 9 Nf κb Protein Ripa Sds page Vimentin Wnt5a β catenin

Corresponding Organization :

Other organizations : Kaohsiung Medical University, National Sun Yat-sen University, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University, E-Da Hospital

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Variable analysis

independent variables

Cellular protein lysates were prepared by homogenization with 1× RIPA lysis buffer (Cell Signaling Technology, Billerica, MA)

dependent variables

Protein concentrations were measured using a protein assay dye (Bio-Rad Laboratories, Hercules, CA)
SDS-PAGE and immunoblotting analysis were performed to detect the expression of the following proteins: E-cadherin, vimentin, α-SMA, fibronectin, WNT5A, β-catenin, NF-κB, and MMP-9

control variables

GAPDH was used as a loading control

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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