Protocol detail

Western Blot Analysis of Inhibitor-Treated Cells

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Western blot analysis of the inhibitor-treated cells was performed using a standardized protocol [35 (link)]. The primary human antibodies used in these analyses included c-MYC, SMO, and β-Actin (Santacruz, CA), AKT, phospho-AKT, S6K, phospho-S6K, GLI1 and SOX2 (Cell Signaling Technology, MA) and, cyclin D1, Bcl-2 and CD133 (BD Biosciences, CA). Immunoreactivity was detected using appropriate peroxidase-conjugated secondary antibodies (Santacruz, CA) and visualized using an enhanced chemiluminescence (ECL) detection system (Pierce, IL).

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Chaturvedi N.K., Kling M.J., Coulter D.W., McGuire T.R., Ray S., Kesherwani V., Joshi S.S, & Sharp J.G. (2018). Improved therapy for medulloblastoma: targeting hedgehog and PI3K-mTOR signaling pathways in combination with chemotherapy. Oncotarget, 9(24), 16619-16633.

Publication 2018

c-MYC β-Actin phospho-AKT phospho-S6K cyclin D1 Bcl-2 CD133 peroxidase-conjugated secondary antibodies

Actin Antibodies Bcl 2 C myc Cells Chemiluminescence Cyclin d1 Human Peroxidase Sox2 Western blot analysis

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Variable analysis

independent variables

Treatment with inhibitor

dependent variables

Protein expression levels of c-MYC, SMO, AKT, phospho-AKT, S6K, phospho-S6K, GLI1, SOX2, cyclin D1, Bcl-2, and CD133

control variables

β-Actin (used as a loading control)

controls

Positive controls: Not explicitly mentioned
Negative controls: Not explicitly mentioned

Annotations

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