Cells were washed with cold PBS, scraped into RIPA buffer and centrifuged. The cell lysates were subjected to 10% SDS-PAGE and transferred to a polyvinylidene fluoride (PVDF) hybridization transfer membrane. The primary antibodies used were as follows: CD44 (ProteinTech group), EGFR (Santa Cruz Laboratories), Snail (Cell Signaling Technology), p-EGFR (Santa Cruz Laboratories), p-AKT (Cell Signaling Technology), p-ERK (Santa Cruz Laboratories), KLF4 (Santa Cruz Laboratories), c-Myc (Santa Cruz Laboratories), Slug (Cell Signaling Technology), Snail (Cell Signaling Technology), Vinculin (Santa Cruz Laboratories). Secondary staining and detection were carried out in accordance with standard protocols (23 (link)).
Protocol detail
Western Blot Analysis of EMT Markers
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Antibodies
Buffer
C myc
Cd44
Cell
Cold
Egfr
Hybridization
Klf4
Membrane
Polyvinylidene fluoride
Ripa
Sds page
Slug
Snail
Vinculin
Corresponding Organization :
Other organizations : Huazhong University of Science and Technology, Tongji Hospital, Pingdingshan University, The Barbara Ann Karmanos Cancer Institute, Wayne State University
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Variable analysis
independent variables
- None explicitly mentioned
- CD44 expression
- EGFR expression
- Snail expression
- P-EGFR expression
- P-AKT expression
- P-ERK expression
- KLF4 expression
- C-Myc expression
- Slug expression
- Snail expression
- Vinculin expression
- None explicitly mentioned
- Positive controls: None mentioned
- Negative controls: None mentioned
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