MED15 is a novel mediator of gene expression programs of chronic inflammation and epithelial–mesenchymal transition in head and neck squamous cell carcinoma - Full Western blots
Description
This dataset includes the full unedited Western blots from the manuscript "MED15 is a novel mediator of gene expression programs of chronic inflammation and epithelial–mesenchymal transition in head and neck squamous cell carcinoma".
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Steps to reproduce
Protein extracts were prepared according to a previously published protocol 71. Briefly, 5 L of protein lysates were loaded onto SDS-polyacrylamide gels and transferred to Immun-Blot PVDF membranes (Bio-Rad Laboratories). After blocking in 5% milk, the membranes were incubated in primary antibodies against the following: MED15 (11566-1-AP, Proteintech, 1:10,000), MED15 (Atlas Antibodies, 1:5,000), MED26 (Cell Signaling Technology, 1:5000), MED27 (CRSP34, Santa Cruz Biotechnology, 1:5000), MED30 (Proteintech, 1:10,000), p63 (4A4, 1:20,000), FRA1 (D80B4, Cell Signaling Technology, 1:5,000), fibronectin (Sino Biological, 1;10,000), vimentin (Cell Signaling Technology, 1:5,000), N-cadherin (BD Transduction Laboratories, 1:5,000), and E-cadherin (Cell Signaling Technology, 1:5,000).The GAPDH MAB374 antibody (EMD Millipore) or -tubulin MAB5564 antibody (EMD Millipore) was used for loading controls at 1:20,000 dilution. Blots were incubated in HRP-conjugated secondary antibodies corresponding to the primary antibody host. Unbound antibodies were washed off in 0.05% Tween-20 in Tris-buffered saline. The LumiGLO peroxidase chemiluminescent substrate kit (SeraCare) was used to detect antibody-labeled proteins, and membranes were imaged using the Bio-Rad ChemiDoc imaging system.