Immunoblot Analysis of Apoptosis Regulators

Immunoblots were carried out as described by our group [23 ,24 (link)]. Original immunoblots see File S1. Antibodies used for this assay were: BCL-XL (#ab32370), survivin (#ab134170), p21 (#ab109520), BAX (#ab32503), BAK (#ab32371), BIM (#ab32158), HDAC3 (#ab32369), GAPDH (#ab128915) from Abcam, Cambridge, UK; MCL-1 (#sc-819), HDAC8 (#sc-374180), HSP90 (#sc-13119), vinculin (#sc-73614) from Santa Cruz Biotechnology, Heidelberg, Germany; cleaved caspase-3 (#cs9661), PARP1 (#cs9542), BID (#cs2002), HDAC1 (#cs34589), HDAC2 (#cs5113), histone H3 (#cs14269), ac-histone H3 (K9) (#cs9649), ac-histone H3 (K18) (#cs9675), ac-histone H3 (K27) (#cs8173) from Cell Signaling, Leiden, The Netherlands; ac-tubulin (#T7451) from Sigma-Aldrich, Taufkirchen, Germany; ac-histone H3 (#06-599) from Merck Millipore, Burlington, MA, USA; and NOXA (#ALX-804-408) from Enzo Life Sciences, New York, NY, USA. HSP90, GAPDH, and vinculin served as independent housekeeping proteins to normalize protein loading. The protein ladders used were the PageRuler^TM pre-stained protein ladder (#26616) and the PageRuler^TM Plus pre-stained protein ladder (#26619) from Thermo Fischer Scientific, Waltham, MA, USA.

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Ashry R., Mustafa A.H., Hausmann K., Linnebacher M., Strand S., Sippl W., Wirth M, & Krämer O.H. (2023). NOXA Accentuates Apoptosis Induction by a Novel Histone Deacetylase Inhibitor. Cancers, 15(14), 3650.

Publication 2023

BCL-XL survivin BAX HDAC3 GAPDH MCL-1 HDAC8 HSP90 vinculin cleaved caspase-3 PARP1 HDAC1 HDAC2 histone H3 ac-tubulin PageRulerTM pre-stained protein ladder PageRulerTM Plus pre-stained protein ladder

Antibodies Assay Caspase 3 Gapdh Histone h3 Hsp90 Immunoblots Parp1 Protein Survivin Tubulin Vinculin

Corresponding Organization :

Other organizations : Mansoura University, Johannes Gutenberg University Mainz, Aswan University, Martin Luther University Halle-Wittenberg, University of Rostock, University Medical Center of the Johannes Gutenberg University Mainz, Humboldt-Universität zu Berlin, Freie Universität Berlin, Universitätsmedizin Göttingen, German Cancer Research Center, Heidelberg University, Charité - Universitätsmedizin Berlin, University of Göttingen

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Variable analysis

independent variables

Not explicitly mentioned

dependent variables

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control variables

HSP90
GAPDH
Vinculin

controls

PageRuler™ pre-stained protein ladder (#26616) and the PageRuler™ Plus pre-stained protein ladder (#26619) from Thermo Fischer Scientific, Waltham, MA, USA

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