Immunoblot Analysis of Protein Expression

Immunoblots were performed following standard procedures^{24 (link)}. Specific antibodies against HDAC6 (7558 S Cell Signalling), acetyl-α-tubulin (ab24610, Abcam), α-tubulin (ab52866, Abcam), acetyl-histone H3-lys9 (9649, Cell Signaling), histone H3 (NB500-171, Novus), PARP (ab32064, Abcam), BMI-1 (05-637, Millipore), SOX2 (AB5603, Millipore), SOX9 (AB5535, Millipore), and β-actin (A5441, Sigma) were used in the study. For secondary antibodies, horseradish peroxidase (HRP)-linked anti-rabbit (7074S, Cell Signalling) or anti-mouse (7076S, Cell Signalling) were used. Detection was performed by chemiluminiscence using NOVEX ECL Chemi Substrate (ThermoFisher).

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Auzmendi-Iriarte J., Saenz-Antoñanzas A., Mikelez-Alonso I., Carrasco-Garcia E., Tellaetxe-Abete M., Lawrie C.H., Sampron N., Cortajarena A.L, & Matheu A. (2020). Characterization of a new small-molecule inhibitor of HDAC6 in glioblastoma. Cell Death & Disease, 11(6), 417.

Publication 2020

ab24610 ab52866 NB500-171 ab32064 AB5603 AB5535 A5441 NOVEX ECL Chemi Substrate

Actin Antibodies Histone h3 Horseradish peroxidase Immunoblots Mouse Novus Rabbit Sox2 Sox9 α tubulin

Corresponding Organization :

Other organizations : Biogipuzkoa Health Research Institute, CIC biomaGUNE, Instituto de Salud Carlos III

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Variable analysis

independent variables

Specific antibodies against HDAC6, acetyl-α-tubulin, α-tubulin, acetyl-histone H3-lys9, histone H3, PARP, BMI-1, SOX2, SOX9, and β-actin

dependent variables

Protein expression levels of HDAC6, acetyl-α-tubulin, α-tubulin, acetyl-histone H3-lys9, histone H3, PARP, BMI-1, SOX2, SOX9, and β-actin

control variables

Standard procedures for immunoblots as described in reference 24

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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