Protein Expression Analysis in Cell Types

Whole cell protein and nuclear protein preparation, as well as western blotting, were performed as we described previously [24 (link), 25 (link)]. For whole protein, the membrane was probed with primary antibodies of anti-E-cadherin (rabbit polyclonal, R&D System, 1:1000), anti-α-smooth muscle actin (SMA) (rabbit polyclonal, R&D System, 1:1000), anti-fibroblast specific protein (FSP)-1 (rabbit polyclonal, Abcam, 1:1000), anti-collagen I (rabbit polyclonal, Calbiochem, 1:1000) and anti-MCP-1 (rabbit polyclonal, Abcam, 1:500); for nuclear protein, the membrane was probed with anti-NF-κB-p65 subunit antibody (rabbit polyclonal, Abcam, 1:1000). The intensities of the blots were determined using an imaging analysis program (Image J, free download from http://rsbweb.nih.gov/ij/). The β-actin was used as internal control. The normalized values in different groups were averaged and expressed as fold change with the mean value of control group as 1.

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Hu J., Zhu Q., Li P.L., Wang W., Yi F, & Li N. (2015). Stem Cell Conditioned Culture Media Attenuated Albumin-Induced Epithelial– Mesenchymal Transition in Renal Tubular Cells. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 35(5), 1719-1728.

Publication 2015

anti-E-cadherin anti-α-smooth muscle actin (SMA) anti-MCP-1 anti-NF-κB-p65 subunit antibody

Actin Anti antibodies Anti antibody Cadherin Cell Collagen i Fibroblast specific protein 1 Mcp 1 Membrane Nf κb p65 subunit Nuclear protein Protein Rabbit Smooth muscle

Corresponding Organization :

Other organizations : Virginia Commonwealth University, Shandong University

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Variable analysis

independent variables

Whole cell protein and nuclear protein preparation

dependent variables

Expression levels of E-cadherin, α-smooth muscle actin (SMA), fibroblast specific protein (FSP)-1, collagen I, and MCP-1 (whole protein)
Expression level of NF-κB-p65 subunit (nuclear protein)

control variables

β-actin as internal control

positive controls

Not specified

negative controls

Not specified

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