Total RNA extraction was carried out with trizol reagent (Invitrogen), and quantitative reverse transcription-PCR (Q-RT–PCR) was performed as described.34 (link)Total protein extracts were obtained and separated as described.34 (link) The following MoAbs were used: mouse anti-MYCN and mouse anti-β-tubulin (Santa Cruz Biotechnology, Santa Cruz, CA, USA); and rat anti-Gal-3 purified monoclonal antibody (Space Import & Export). Immunoreactive bands were visualized by enhanced chemoluminescence (Perkin Elmer, Waltham, MA, USA).
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Quantitative Analysis of Protein Expression
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Other organizations : Sapienza University of Rome, CTO Andrea Alesini, National Agency for New Technologies, Energy and Sustainable Economic Development, Bambino Gesù Children's Hospital, Istituti di Ricovero e Cura a Carattere Scientifico, Alder Hey Children's NHS Foundation Trust
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Variable analysis
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- Total RNA extraction with trizol reagent (Invitrogen)
- Quantitative reverse transcription-PCR (Q-RT–PCR) as described
- Total protein extracts obtained and separated as described
- Immunoreactive bands visualized by enhanced chemoluminescence (Perkin Elmer, Waltham, MA, USA)
- Mouse anti-MYCN
- Mouse anti-β-tubulin (Santa Cruz Biotechnology, Santa Cruz, CA, USA)
- Rat anti-Gal-3 purified monoclonal antibody (Space Import & Export)
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