Journal: European journal of pharmacology

Article Title: The opioid antagonist, β-funaltrexamine, inhibits NF-κB signaling and chemokine expression in human astrocytes and in mice

doi: 10.1016/j.ejphar.2015.05.040

Figure Lengend Snippet: β-FNA actions are downstream of IL-1R. NHA were exposed to human recombinant IL-1β (3 ng/ml) for 0.5 or 3 h, followed by washout of stimulus. Cells were then exposed to β-FNA (10 μM) for 21 h. CXCL10 protein levels in the media then were measured by ELISA. The data represent mean ± S.E.M. (n=5–14) and are presented as percent control (24 h IL-1β stimulation). *P < 0.05 vs. 3 h control.

Article Snippet: 2.3 CXCL10 protein and RNA expression A standard dual-antibody solid phase immunoassay (ELISA Development Kit; Peprotech) was used for quantitation of secreted CXCL10 in NHA cell supernatants, and in mouse plasma and whole brain homogenates, according to the manufacturer’s instructions and as previously described ( Davis et al., 2007 ).

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Control

Journal: European journal of pharmacology

Article Title: The opioid antagonist, β-funaltrexamine, inhibits NF-κB signaling and chemokine expression in human astrocytes and in mice

doi: 10.1016/j.ejphar.2015.05.040

Figure Lengend Snippet: Inhibitory effects of β-FNA persist after drug washout. NHA were unstimulated, exposed to human recombinant IL-1β (3 ng/ml) alone or exposed to IL-1β + β-FNA (10 μM) for 24 h. In another treatment group, cells were pre-treated with β-FNA for 60 min. followed by drug washout. Cells were then exposed to IL-1β for 24 h. CXCL10 protein levels in the media then were measured by ELISA. The data represent mean ± S.E.M (n=11–12). *P < 0.05 vs. IL-1β alone.

Article Snippet: 2.3 CXCL10 protein and RNA expression A standard dual-antibody solid phase immunoassay (ELISA Development Kit; Peprotech) was used for quantitation of secreted CXCL10 in NHA cell supernatants, and in mouse plasma and whole brain homogenates, according to the manufacturer’s instructions and as previously described ( Davis et al., 2007 ).

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay

Journal: European journal of pharmacology

Article Title: The opioid antagonist, β-funaltrexamine, inhibits NF-κB signaling and chemokine expression in human astrocytes and in mice

doi: 10.1016/j.ejphar.2015.05.040

Figure Lengend Snippet: Naltrexone does not inhibit IL-1β-induced CXCL10 expression in NHA. NHA were exposed to IL-1β (3 ng/ml) alone or co-exposed to IL-1β and β-FNA (10 μM) or naltrexone (10 μM) for 24 h. CXCL10 protein levels in the media were then measured by ELISA. Data represent mean ±S.E.M. (n=9–12). *P < 0.05 vs. IL-1β alone.

Article Snippet: 2.3 CXCL10 protein and RNA expression A standard dual-antibody solid phase immunoassay (ELISA Development Kit; Peprotech) was used for quantitation of secreted CXCL10 in NHA cell supernatants, and in mouse plasma and whole brain homogenates, according to the manufacturer’s instructions and as previously described ( Davis et al., 2007 ).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: European journal of pharmacology

Article Title: The opioid antagonist, β-funaltrexamine, inhibits NF-κB signaling and chemokine expression in human astrocytes and in mice

doi: 10.1016/j.ejphar.2015.05.040

Figure Lengend Snippet: β-FNA inhibits IL-1β-induced CXCL10 expression in NHA. A) NHA were co-exposed to human recombinant IL-1β (3 ng/ml) and β-FNA (0.3–100μM) for 24 h. CXCL10 protein levels in the media were then measured by ELISA. The data represent mean + S.E.M (n=11–15). B) NHA were treated with IL-1β (3 ng/ml) alone or co-exposed to IL-1β and β-FNA (10 μM) for 8 h. Total RNA was isolated and CXCL10 and GAPDH mRNA assessed by real-time PCR. The data represent mean ± S.E.M. (n=5–6). *P < 0.05.

Article Snippet: 2.3 CXCL10 protein and RNA expression A standard dual-antibody solid phase immunoassay (ELISA Development Kit; Peprotech) was used for quantitation of secreted CXCL10 in NHA cell supernatants, and in mouse plasma and whole brain homogenates, according to the manufacturer’s instructions and as previously described ( Davis et al., 2007 ).

Techniques: Expressing, Recombinant, Enzyme-linked Immunosorbent Assay, Isolation, Real-time Polymerase Chain Reaction

Journal: European journal of pharmacology

Article Title: The opioid antagonist, β-funaltrexamine, inhibits NF-κB signaling and chemokine expression in human astrocytes and in mice

doi: 10.1016/j.ejphar.2015.05.040

Figure Lengend Snippet: Inhibition of the ubiquitination pathway is likely involved in the anti-inflammatory actions of β-FNA. NHA were exposed to human recombinant IL-1β (3 ng/ml) alone, IL-1β + β-FNA (5 μM), IL-1β + PYR41 (4 nM; inhibitor of Ub-activating enzyme E1), or, IL-1β + β-FNA + PYR41 for 24 h. CXCL10 protein levels in the media then were measured by ELISA. The data represent mean ± S.E.M. (n=6). **P< 0.01, ***P < 0.001 vs. IL-1β + β-FNA.

Article Snippet: 2.3 CXCL10 protein and RNA expression A standard dual-antibody solid phase immunoassay (ELISA Development Kit; Peprotech) was used for quantitation of secreted CXCL10 in NHA cell supernatants, and in mouse plasma and whole brain homogenates, according to the manufacturer’s instructions and as previously described ( Davis et al., 2007 ).

Techniques: Inhibition, Ubiquitin Proteomics, Recombinant, Enzyme-linked Immunosorbent Assay

Journal: European journal of pharmacology

Article Title: The opioid antagonist, β-funaltrexamine, inhibits NF-κB signaling and chemokine expression in human astrocytes and in mice

doi: 10.1016/j.ejphar.2015.05.040

Figure Lengend Snippet: β-FNA inhibits LPS-induced CXCL10 protein levels in brain, but not plasma. C57BL/6J mice were injected (i.p.) with saline (n = 7), LPS (5 mg/kg; n = 5), β-FNA (28 mg/kg; n = 3), or LPS + β-FNA (n = 5). At 6 h post-injections, plasma and whole brain were collected on ice. CXCL10 protein levels in plasma and whole brain tissue homogenates were determined by ELISA. Data represent the mean ± S.E.M.; *P < 0.05 vs. LPS.

Article Snippet: 2.3 CXCL10 protein and RNA expression A standard dual-antibody solid phase immunoassay (ELISA Development Kit; Peprotech) was used for quantitation of secreted CXCL10 in NHA cell supernatants, and in mouse plasma and whole brain homogenates, according to the manufacturer’s instructions and as previously described ( Davis et al., 2007 ).

Techniques: Clinical Proteomics, Injection, Saline, Enzyme-linked Immunosorbent Assay