Protein Extraction and Immunoblotting Protocol

Cells were lysed using the ELB lysis buffer containing 150 mM NaCl, 50 mM Hepes pH 7.5, 5 mM EDTA, 0.1% NP-40, 20 mM β-glycerophosphate, 0.5 mM sodium orthovanadate and a protease inhibitor (Roche). The immunoblotting procedure was performed as described [38 (link)]. The antibodies used in this study were directed against MCPyV-LT (CM2B4; Santa Cruz Biotechnologies), β-tubulin (TUB 2.1; Sigma-Aldrich, Ottobrunn, Germany) and vinculin (hVIN-1; Sigma-Aldrich). Uncropped blots are given in Supplementary Figures S9 and S10)

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Sarma B., Willmes C., Angerer L., Adam C., Becker J.C., Kervarrec T., Schrama D, & Houben R. (2020). Artesunate Affects T Antigen Expression and Survival of Virus-Positive Merkel Cell Carcinoma. Cancers, 12(4), 919.

Publication 2020

CM2B4 TUB 2.1 hVIN-1

Antibodies Buffer Cells Edta Hepes Mcpyv Nacl Np 40 Orthovanadate Protease inhibitor Sodium Tubulin Vinculin β glycerophosphate

Corresponding Organization :

Other organizations : Universitätsklinikum Würzburg, Essen University Hospital, German Cancer Research Center, Heidelberg University, Centre Hospitalier Universitaire de Tours

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Variable analysis

independent variables

Lysis buffer composition: 150 mM NaCl, 50 mM Hepes pH 7.5, 5 mM EDTA, 0.1% NP-40, 20 mM β-glycerophosphate, 0.5 mM sodium orthovanadate, protease inhibitor (Roche)

dependent variables

Protein expression levels of MCPyV-LT, β-tubulin, and vinculin (measured by immunoblotting)

control variables

Immunoblotting procedure (described in reference 38)

positive controls

Not specified

negative controls

Not specified

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