More detailed information is available as Supp Data. In brief; MTS and clonogenicity assays were performed as previously described (26 ). Western blot analyses were performed by standard methods (26 ). ELISA HGF and VEGF levels were measured in MPNST cell conditioned media using enzyme-linked immunosorbent assay (ELISA). The assays were constructed and preformed following manufacturer’s instructions (R&D, Minneapolis, MN). Migration and invasion assays were conducted using modified Boyden chambers as previously described (27 (link)). qRTPCR for MMP2 was conducted as previously described (28 (link)).MET gene sequencing procedure and primers as well as siRNA and shRNA transfection and transduction procedures are detailed in Supp data.
Protocol detail
Characterization of MPNST Cell Signaling
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Assays
Cell
Conditioned media
Enzyme linked immunosorbent assay
Gene procedure
Mmp2
Mpnst
Primers
Shrna
Sirna
Transfection
Vegf
Western blot analyses
Corresponding Organization : The University of Texas MD Anderson Cancer Center
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Variable analysis
independent variables
- MET gene sequencing procedure and primers
- SiRNA and shRNA transfection and transduction procedures
- MTS and clonogenicity
- HGF and VEGF levels in MPNST cell conditioned media
- Migration and invasion
- MMP2 expression
- Western blot analyses were performed by standard methods
- Positive control: Not specified
- Negative control: Not specified
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