Standard Western blotting was carried as described previously (7 (link)). PARP, caspase-8, caspase-3, TRAIL-R2, and Bid antibodies were purchased from Cell Signaling Technology, cIAP1 from Enzo, FADD from BD Biosciences, FLIP antibody from AdipoGen, and β-actin from Sigma. For absolute quantification of FLIP and caspase-8, images were captured using an Odyssey Imaging System (LICOR) at 12-bit dynamic range. Quantification of protein expression amounts was conducted as described previously using recombinant proteins and a HeLa cell line standard (8 (link)).
Grinkevitch V., Wappett M., Crawford N., Price S., Lees A., McCann C., McAllister K., Prehn J., Young J., Bateson J., Gallagher L., Michaut M., Iyer V., Chatzipli A., Barthorpe S., Ciznadija D., Sloma I., Wesa A., Tice D.A., Wessels L., Garnett M., Longley D.B., McDermott U, & McDade S.S. (2022). Functional Genomic Identification of Predictors of Sensitivity and Mechanisms of Resistance to Multivalent Second-Generation TRAIL-R2 Agonists. Molecular Cancer Therapeutics, 21(4), 594-606.
Corresponding Organization : Queen's University Belfast
Other organizations :
Wellcome Sanger Institute, Royal College of Surgeons in Ireland, The Netherlands Cancer Institute, Champions Oncology (United States), AstraZeneca (United States), Delft University of Technology
Protein expression levels of PARP, caspase-8, caspase-3, TRAIL-R2, Bid, cIAP1, FADD, FLIP, and β-actin
control variables
HeLa cell line
Recombinant proteins for FLIP and caspase-8 quantification
positive controls
Not explicitly mentioned
negative controls
Not explicitly mentioned
Annotations
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