Western Blot Analysis of Cell Signaling

WB was performed as described [49 (link)]. Antibodies used were the following: CK1α, PARP, Mcl1, Ser45 β-catenin, Ser33/37/Thr41 β-catenin, total β-catenin, Ser473 AKT, total AKT (Cell signaling Technology, MA, USA); Mdm2 (Millipore, Italy), GAPDH (Ambion, USA); α-tubulin (Sigma-Aldrich, Italy); Bak (Merck, MA, USA); Bax, p21, p53 (Becton Dickinson, Italy); Caspase 3 (Enzo Life Science, UK). Images were acquired using the Image Quant LAS 500 chemiluminescence detection system (GE Healthcare, USA). Densitometric analysis was performed with Quantity One software (Bio-Rad, Italy).

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Manni S., Carrino M., Manzoni M., Gianesin K., Nunes S.C., Costacurta M., Tubi L.Q., Macaccaro P., Taiana E., Cabrelle A., Barilà G., Martines A., Zambello R., Bonaldi L., Trentin L., Neri A., Semenzato G, & Piazza F. (2017). Inactivation of CK1α in multiple myeloma empowers drug cytotoxicity by affecting AKT and β-catenin survival signaling pathways. Oncotarget, 8(9), 14604-14619.

Publication 2017

Mcl1 Ser45 β-catenin Ser33/37/Thr41 β-catenin total β-catenin Ser473 AKT total AKT GAPDH α-tubulin Caspase 3 Image Quant LAS 500 chemiluminescence detection system Quantity One software

Antibodies Caspase 3 Chemiluminescence Ck1α Densitometric Gapdh Mcl1 Mdm2 α tubulin β catenin

Corresponding Organization :

Other organizations : University of Padua, Veneto Institute of Molecular Medicine, University of Milan, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Istituto Oncologico Veneto, Istituti di Ricovero e Cura a Carattere Scientifico

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Variable analysis

independent variables

Antibodies used: CK1α, PARP, Mcl1, Ser45 β-catenin, Ser33/37/Thr41 β-catenin, total β-catenin, Ser473 AKT, total AKT, Mdm2, GAPDH, α-tubulin, Bak, Bax, p21, p53, Caspase 3

dependent variables

Protein expression levels of CK1α, PARP, Mcl1, Ser45 β-catenin, Ser33/37/Thr41 β-catenin, total β-catenin, Ser473 AKT, total AKT, Mdm2, GAPDH, α-tubulin, Bak, Bax, p21, p53, Caspase 3

control variables

Western blotting (WB) protocol as described in reference [49]
Chemiluminescence detection system (Image Quant LAS 500)
Densitometric analysis software (Quantity One)

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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