Western blotting was conducted as in past reports.25 (link),26 (link) Briefly, after proteins were transferred to PVDF membranes, these blots were probed using primary antibodies specific for SOX2 (Seven Hills, OH, USA), β-actin (Beyotime, Nantong, China), LLGL2 (Abcam, MA, USA) or E-cadherin (Cell Signaling Technology, MA, USA). HRP-linked secondary antibodies specific for mouse-IgG (ZSGB-Bio, Beijing, China) or rabbit IgG (Abcam) were then used for protein detection, β-actin was used as a control for normalization, and Image-Pro Plus 6.0 was used for quantification.
Protocol detail
Western Blotting of Key Proteins
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Corresponding Organization : Xiamen University
Other organizations : University of Edinburgh, MRC Centre for Regenerative Medicine, Zhoushan Hospital, University of Rochester
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Variable analysis
independent variables
- None explicitly mentioned
- Protein expression levels of SOX2, β-actin, LLGL2, and E-cadherin
- β-actin protein expression used for normalization
- Positive control: β-actin
- Negative controls: not explicitly mentioned
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