BDNF Antibody - #DF6387

Fig. 4. Ift172 haploinsufficiency led to a dysfunctional BDNF-TrkB signaling pathway and reduced synaptic density. (A) Compared with WT littermates, Ift172+/- mice showed reduced expression of BDNF, p-TrkB, p-Erk1/2, and PSD95 in the cortex. (B) Quantification of relative protein expressing levels (n = 3 per group). (C) Immunohistochemical staining showed decreased BDNF expression in the brain of Ift172+/- mice compared with WT littermates. Scale bars = 2 mm (left column) and 10 μm (right column). (D) Immunofluorescent staining of Ift172 and BDNF in the cortex of WT and Ift172+/- mice. Scale bar = 20 µm. (E) Quantification analysis of the expression of Ift172 and BDNF proteins (n = 5 per group). (F) Schematic diagram showed the construct of an adeno-associated virus and the strategy of sparse labeling. (G) Confocal images of representative pyramidal neurons and their apical dendrites in the CA1 region of WT and Ift172+/- mice. Scale bars = 10 μm (top row) and 5 μm (bottom row). (H) Compared with WT littermates, Ift172+/- mice presented significantly less spine density. (WT: n = 11 dendrites from 3 mice; Ift172+/-: n = 15 dendrites from 3 mice). (I) Representative electron micrographs of the synaptic structures in the CA1 area. Red arrows indicated synapses. Scale bar = 1 μm. (J) Quantitative analysis of the synaptic density in WT and Ift172+/- mice. (n = 5 per group). Data were presented as the mean ± s.e.m. (*p < 0.05, **p < 0.01, and ***p < 0.001 with Student’s t-test). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Figure 3—Exogenous MANF promoted corneal nerve regeneration and BDNF and NGF expression. Normal and diabetic corneas pretreated through subconjunctival injection with PBS (as the control) or rhMANF 24 h before, 0 h, and 24 h after removal of the corneal epithelium. At 48 h, corneal epithelial cells were collected for Western blotting. At 5 days postwounding, corneal whole-mount staining was performed to observe nerve regeneration. Corneal sensation was tested before epithelium debridement until 7 days postwounding. To observe nerve regeneration and corneal sensation, another injection was performed at 48 h postwounding. A: Staining for b-tubulin III was performed, and images of the entire cornea were captured as shown in the top panels, with central cornea images shown in the bottom panels. B: Nerve densities of entire corneas (A, top panels) were calculated from the areas staining positive for b-tubulin III using Image J software (n 5 3). C: Corneal sensation is represented with a line chart (n 5 5). The time immediately before epithelium debridement was defined as 0 h. D: Western blot bands observed with or without rhMANF. Two samples/lane represent each condition, three corneas as one sample. E: Western blot analysis of BDNF expression (n 5 4 samples). F: Western blot analysis of NGF expression (n 5 4 samples). *P , 0.05, **P , 0.01, ***P , 0.001 (one-way ANOVA). DM, wounded mice with diabetes mellitus; DM 1 MANF, wounded mice with diabetes mellitus treated with rhMANF; NL, wounded normoglycemic mice treated with PBS; NL 1 MANF, wounded normoglycemic mice treated with rhMANF; n.s., not significant.

Figure 3 Gene and protein verification in sciatic nerve injury in rats treated with AKBA. (A) mRNA expression levels (relative to GAPDH) of NGFR (A1), NGF (A2), BDNF (A3), and MPO (A4) in the rat sciatic nerve using qPCR. (B) Protein expression levels (relative to GAPDH) of NGFR (B2), NGF (B3), BDNF (B4), and MPO (B5) in the rat sciatic nerve by western blot assay. Data are expressed as the mean ± SD (n = 3 rats/group) and were analyzed using a one-way analysis of variance followed by the least significant difference test. BDNF: Brain derived neurotrophic factor; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; MPO: myeloperoxidase; NGF: nerve growth factor; NGFR: nerve growth factor receptor; qPCR: real-time polymerase chain reaction.

Figure 6.| ZL activated cAMP-CREB-BDNF signaling. (A–C) Protein expressions of BDNF, p-CREB and CREB in the hippocampus at the end of experiment determined by western blotting.Quantified protein level was normalized to that of GAPDH using densitometry (n = 3)

Fig. 6. |Expression of AKT/FOXO1 signaling pathway (A); mitochondrial function markers(B); BDNF signaling-related proteins (C); and representative confocal microscopy images of BDNF staining (green, magnification: ×40,scale Bar 50 μm) (D: a, 8 M–SED; b, 26 M–SED;c, 18 M–MICT; d, 8 M–MICT); and serum BDNF levels (E); and correlation analysis (F).

Fig. 6. |Expression of AKT/FOXO1 signaling pathway (A); mitochondrial function markers(B); BDNF signaling-related proteins (C); and representative confocal microscopy images of BDNF staining (green, magnification: ×40,scale Bar 50 μm) (D: a, 8 M–SED; b, 26 M–SED;c, 18 M–MICT; d, 8 M–MICT); and serum BDNF levels (E); and correlation analysis (F).