Immunoblotting analysis of epigenetic and apoptotic markers

Floating and attached cells were lysed in Radioimmunoprecipitation assay (RIPA) buffer. Protein concentration was determined by using Micro BCA TM protein assay kit (Thermo Fisher). Immunoblotting was performed as described previously (5 (link)). The following antibodies were used to detect G9a (ab185050, Abcam, RRID:AB_2792982), cPARP (ab32064, Abcam, RRID:AB_777102), MYCN (B8.48, Santa Cruz, SC-53993, RRID:AB_831602), cCaspase 3 (9664, Cell Signaling Technology, RRID:AB_2070042), LC3B (L7543, Sigma, RRID:AB_796155), histone H3 (ab10799, Abcam, RRID:AB_470239), dimethyl K9 histone H3 (ab1220, Abcam, RRID:AB_449854), and β-Actin (A3854, Sigma, RRID:AB_262011), according to manufacturer's instructions.

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Bellamy J., Szemes M., Melegh Z., Dallosso A., Kollareddy M., Catchpoole D, & Malik K. (2020). Increased Efficacy of Histone Methyltransferase G9a Inhibitors Against MYCN-Amplified Neuroblastoma. Frontiers in Oncology, 10, 818.

Publication 2020

Micro BCA TM protein assay kit ab185050 ab32064 L7543 ab10799 ab1220 A3854

Actin Antibodies Assay Buffer Cells Histone h3 Mycn Protein Radioimmunoprecipitation assay

Corresponding Organization : University of Bristol

Other organizations : Southmead Hospital, Children's Hospital at Westmead

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Protein expression levels of G9a, cPARP, MYCN, cCaspase 3, LC3B, histone H3, dimethyl K9 histone H3, and β-Actin

control variables

None explicitly mentioned

controls

Positive control: None mentioned
Negative control: None mentioned

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