Fig 1: Brain-derived neurotrophic factor (BDNF) activation of mTORC1 in hippocampal neurons is palmitoylation-dependent. (A) Scheme of the BDNF stimulation paradigm. DIV14–16 primary rat hippocampal neurons were placed in artificial cerebrospinal fluid (ACSF) for 2 h before stimulation with 100 ng/mL BDNF or 5 mM Leu. Fifteen minutes prior to stimulation, cells were treated with ethanol (EtOH) or 50 μM 2BP to inhibit palmitoylation. Neurons were stimulated for 30 min with BDNF or Leu. (B) Lysates were blotted to detect p-S6 (top), S6 total (middle), and tubulin (bottom) levels. Right panel histogram shows quantified data from multiple experiments, normalized to the regular growth media with B27 (+B27) condition [two-way ANOVA: 2BP p < 0.0001 (F(2) = 28.41), treatment p < 0.0001 (F(1) = 59.31), interaction p = 0.0017 (F(2) = 9.52); N = 3–5; Bonferroni post hoc test ***p < 0.001]. Panels are composites of the same Western blot image. (C) Neurons were stimulated as in B with BDNF, but 15 min prior to stimulation cells were treated with EtOH or 100 nM rapamycin (Rap) to inhibit mTOR.
Fig 2: BDNF level decreases after SAH. (A) Mortality rate, (B) SAH grade scores, (C) western blot analysis of BDNF protein in basal cortex and (D) ELISA analysis of BDNF concentration in the basal cortex, in the sham and SAH groups at 6, 12, 24, 48 and 72 h. (B) P=0.88; (C and D) P=0.01. *P<0.05 vs. sham; one-way analysis of variance comparison test, n=6 in each group. BDNF, brain-derived neurotrophic factor; SAH, subarachnoid hemorrhage; ns, not significant.
Fig 3: Exogenous BDNF attenuates neurological deficit at 72 h after SAH. (A) Representative brain photos of sham, SAH + vehicle and SAH + BDNF groups (left), SAH grade scores (right), (B) mortality rate, (C) modified Garcia score, (D) beam balance test score and (E) rotarod test score in the sham, SAH + vehicle and SAH + BDNF groups. (n=6, *P<0.05). (A) P=0.73 and (C-E) P=0.01. *P<0.05; one-way analysis of variance comparison test, n=6 in each group. BDNF, brain-derived neurotrophic factor; SAH, subarachnoid hemorrhage.
Fig 4: GDNF stimulated axon outgrowth from DRGs into Matrigel in the 3D explant model. NFG, GDNF, or BDNF was added to the culture media. (A) Immunofluorescent staining of mouse DRG in a Matrigel drop. Axons were visualized with antibody staining against NF200 (red fluorescence), nuclei counterstained with DAPI, and Nestin-positive cells (green fluorescence). Scale bar: 250 µm. (B) A graph presenting the quantification of axon growth. Data are presented as mean ± standard deviation (SD). * p < 0.05, n = 4.
Fig 5: Relative levels of BDNF, AKT, and MAPK following BDNF or BDNF-MSC treatment in MECP2 knockout mice. The activity of the downstream signaling pathways of BDNF in mice was determined using western blotting (A). Phosphorylation of MAPK represents a change in the activities of cell signaling pathways. (B–E) Relative intensity is represented in graphs. *p < 0.05, **p < 0.01, ***p < 0.001.
Supplier Page from Abcam for Recombinant Human/Murine/Rat BDNF protein (Active)