Western Blot Analysis of Neural Markers

Cell lysates were created and Western blotting conducted as described (24 (link)), blocking without detergent. Membranes were probed with antibodies for MAF (Imgenex), SOX9 (Santa Cruz), S100β (Dako), MBP (Chemicon), BLBP (Millipore), DEPTOR (Novus), cleaved-caspase 3, pS6, S6, p4E-BP1, 4E-BP1, p473AKT, AKT, and β-ACTIN (Cell Signaling) as a loading control. Horseradish peroxidase-conjugated secondary antibodies (BioRad) were used with ECL Plus developing system (Amersham Biosciences).

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Brundage M.E., Tandon P., Eaves D.W., Williams J.P., Miller S.J., Hennigan R.H., Jegga A., Cripe T.P, & Ratner N. (2014). MAF mediates crosstalk between Ras-MAPK and mTOR signaling in NF1. Oncogene, 33(49), 5626-5636.

Publication 2014

DEPTOR 4E-BP1 p473AKT β-ACTIN Horseradish peroxidase-conjugated secondary antibodies ECL Plus developing system

4e bp1 Actin Antibodies Caspase 3 Cell Detergent Horseradish peroxidase Membranes Novus S100β Sox9

Corresponding Organization :

Other organizations : University of Cincinnati Medical Center, Cincinnati Children's Hospital Medical Center

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Variable analysis

independent variables

Cell lysate treatment

dependent variables

Protein expression levels of MAF, SOX9, S100β, MBP, BLBP, DEPTOR, cleaved-caspase 3, pS6, S6, p4E-BP1, 4E-BP1, p473AKT, AKT

control variables

Protein loading control (β-ACTIN)

controls

Positive control: Not specified
Negative control: Not specified

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