Cells were cold lysed in an NP40-based lysis buffer while frozen tumor samples were homogenized in lysis buffer using the Roche MagNA Lyser instrument (Roche). Following protein extraction, cellular and tumor lysates were prepared and separated on an SDS-PAGE gel prior transfer and immunoblotting as described previously (25 (link)). Information on primary and secondary antibodies used can be found in Supplemental File 1 , Table 1 .
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Protein Extraction and Immunoblotting
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Corresponding Organization : Georgetown University Medical Center
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Variable analysis
independent variables
- Cell lysis method (cold lysis in NP40-based buffer)
- Tumor sample homogenization method (Roche MagNA Lyser instrument)
- Cellular and tumor lysate protein expression levels (as measured by SDS-PAGE and immunoblotting)
- Protein extraction method
- SDS-PAGE and immunoblotting procedures (as described previously in reference 25)
- Positive controls: Not explicitly mentioned
- Negative controls: Not explicitly mentioned
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