human anti beta ngf antibody  (R&D Systems)

Journal: Journal of Neurochemistry

Article Title: Cerebral Dopamine Neurotrophic Factor ( CDNF ) Acts as a Trophic Factor Promoting Neuritogenesis in the Dorsal Root Ganglia ( DRG ) Neurons Through Activation of the PI3K Signaling Pathway

doi: 10.1111/jnc.70194

Figure Lengend Snippet: CDNF's trophic activity appears to be independent of both NGF and TrkA receptor signaling. Pharmacological inhibitor (GNF‐5837) of Tropomyosin Receptors kinase (+Trki) or anti‐nerve growth factor (NGF) antibody (+α‐NGF) or both (Trki+α‐NGF) were added to dorsal root ganglia (DRG) explants stimulated with NGF 30 ng/mL or cerebral dopamine neurotrophic factor (CDNF, 18 μg/mL). (A) Representative images of DRG explants immunostained with anti‐TuJ‐1 antibody (green) treated with NGF for 72 h (upper images) or with CDNF (lower images) in the absence (control—Ctr) or in the presence of Trki (+Trki—20 μM), anti‐NGF (+α‐NGF—10 μM) or both (Trki + α‐NGF). (B) Neurite length and (C) neuritic area for the conditions displayed in panel A. (D) Cell toxicity assays measured by LDH for the conditions displayed in panel A. Number of Intersections in the presence of NGF (E) or CDNF (F) for the conditions displayed in panel A. Area under the curve (AUC) relative to number of intersections in shown in the inset. The percentage of neurite outgrowth inhibition for each condition is shown at the top in (B). Scale bar = 300 μm. Data are presented as mean ± SEM from n = 3 independent experiments. One‐way ANOVA, with Tukey's post hoc test for significance. * p < 0.05 indicates statistical significance.

Article Snippet: Culture media were then replaced by medium only (deprived group), or media containing NGF (30 ng/mL or as stated in Figure legends), CDNF, or CDNF‐ΔKTEL (both at 1.8 μg/mL or 18 μg/mL or stated in Figure legends) for an additional 48 h. To unravel the signaling pathways involved in neuritogenesis, we incubated DRG cultures with wortmannin (400 nM ; Cat. No. W3144 Merck) or LY294002 (From 4 to 400 nM ); cat. No. S1105 Selleckchem, both are PI3K pathway inhibitors; rapamycin (20 nM ; Cat. No. S‐015 Merck), a mTOR pathway inhibitor; GNF ‐5837 (20 μM ; Cat. No. S7519 Selleckchem), a pan‐Trk inhibitor; and anti‐ NGF antibody (10 μM ; Cat. No. AF256 ‐ NA RD Systems, RRID:AB_2106738) to inhibit NGF binding to its receptor.

Techniques: Activity Assay, Control, Inhibition