Total cell lysates were separated by 8% or 10% SDS-PAGE and then electroblotted and transferred onto PVDF membranes.31 (link) After incubation with primary antibodies specific for COL1A1 (1:1000; sc-293182, Santa Cruz, TX, USA), CD44 (1:2000; #37259, CST, Boston, USA), E-cadherin (1:2000; #14472, CST, Boston, USA), vimentin (1:2000; #5741, CST, Boston, USA), TGFBI(1:1000; 10188–1-AP, proteintech, IL, USA), MMP9 (1:2000; #15561, CST, Boston, USA) and β-actin (1:4000,sc-81178, Santa Cruz, TX, USA) at 4 °C overnight, the blots were incubated with the corresponding secondary antibodies at room temperature, and the proteins were detected with an ECL kit.
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Western Blot Analysis of Extracellular Matrix Proteins
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Corresponding Organization :
Other organizations : Guangzhou Medical University, Third Affiliated Hospital of Guangzhou Medical University, Zhongshan People's Hospital
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Variable analysis
independent variables
- SDS-PAGE gel percentage (8% or 10%)
- Protein expression levels of COL1A1, CD44, E-cadherin, vimentin, TGFBI, and MMP9
- β-actin as a loading control
- No positive or negative controls were explicitly mentioned.
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