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Notes: Sympathetic neurons were grown in the presence of 2.5S NGF prior to injection with cDNAs from the X-chromosome-linked inhibitor of apoptosis gene. (2659)

Notes: The 2.5S Nerve Growth Factor was used to culture rat dorsal root ganglia neurons. The primary rat cells were infected with an adenovirus expressing E. coli β-galactosidase (lacZ) and the enzyme was detected by immunocytochemistry with the Anti-β-Galactosidase mAb. Primary cells infected with either an HSP27 adenovirus or LacZ adenovirus were withdrawn from NGF and survival assayed 48 hours later with the CellTiter 96^® Non-Radioactive Cell Proliferation Assay. Only those cells expressing HSP27 showed some survival and percent survival was dependent upon the multiplicity of infection. (0788)

Notes: To test the efficacy of TrkA, B and C-IgG fusions to their respective ligands, PC12 cells were stimulated Nerve Growth Factor (NGF) and TrkA-IgG, TrkB-IgG, or TrkC-IgG. Only the TrkA-IgG blocked tyrosine phosphorylation as judged by a pan phosphoTrk antibody. The TrkB-IgG blocked the activity of Brain-Derived Neurotrophic Factor (BDNF) on cortical cells and TrkC-IgG blocked Neurotrophin-3 (NT-3) action. Promega's BDNF, NT-3 and NGF, 2.5S, were used. (1452)

Notes: The CytoTox 96^® Non-Radioactive Cytotoxicity Assay was used to assess LDH release from primary cortical neurons following 24hr exposure to glutamate. Various compounds including 2.5S Nerve Growth Factor were assessed for their ability to block the glutamate-induced cytotoxicity. One of the neuroprotective factors, estrogen, was also examined for induction protein tyrosine kinase activity with the the SignaTECT^® Protein Tyrosine Kinase Assay System. The induction of PTK activity peaked at 1min and returned to normal within 10min. (0367)

Notes: 2.5S mNGF was used to differentiate PC12 cells. The differentiated cells were then microinjected with expression plasmids. (0922)

Notes: The SP6 and T7 RNA Polymerases were used to produce RNA probes for Northern Analysis. The authors examined the half-life of the m4 muscarinic receptor in PC12 cells and transformed PC12 cells following treatment with either murine 2.5S NGF, human recombinant basic FGF or human recombinant EGF. PC12 cells were also treated with the three factors, nuclei isolated, and nuclear run-on transcription assay were performed. (0814)

Notes: The 2.5S mNGF was used to differentiate PC12 cells in culture. (1219)

Notes: NGF was used in this study of neurite outgrowth in PC12 cells under a variety of conditions. Protein Kinase C was used to in vitro phosphorylate a bacterially expressed protein and the phospho protein was assayed by gel shift. Many details of the phosphorylation are provided. (0304)

Notes: Superior cervical ganglia were maintained in culture with or without the 2.5S NGF. RNA was isolated from these cultures and used for differential display with RT-PCR. The RT-PCR products were cloned with the pGEM^®-T Vector System. (0998)

Notes: Reporter vectors (produced with the pGL2 Basic Vector) were transiently transfected into PC12 cells and promoter activity was measured in response to 2.5S NGF. Promoter constructs were generated by PCR, subcloned into the pGEM^®-T Vector and sequenced. The pGEM^®-luc Vector was used to generated an antisense luc RNA probe used in RNase protection assays (using RNase ONE™ Ribonuclease) to follow the transcription of luciferase DNA following NGF treatment. The early growth reaction transcription factor and variants were expressed in the TNT^® SP6 Coupled Reticulocyte Lysate System and used to perform gel shifts with promoter elements. The SP1 Consensus Oligonucleotide was used to compete transcription factor binding to the promoter elements. (1493)

Notes: Promega's 2.5S Nerve Growth Factor (NGF; 50ng/ml) was added to the culture medium three days prior to assay to induce PC12 cell differentiation. (0774)