Journal: Neural Regeneration Research

Article Title: Transplantation of human umbilical cord blood mesenchymal stem cells to treat a rat model of traumatic brain injury ★

doi: 10.3969/j.issn.1673-5374.2012.10.004

Figure Lengend Snippet: Neurotrophic factor expression in surrounding injured brain tissues following human umbilical cord blood mesenchymal stem cell transplantation. (A–C) Nerve growth factor (NGF) protein expression (immunohistochemistry, × 200); (D–F) brain-derived neurotrophic factor (BDNF) protein expression (immunohistochemistry, × 200); (G–I) BDNF mRNA expression ( in situ hybridization, × 400). (A, D, G) Results (absorbance) are expressed as mean ± SD from six rats in each group at each time point. a P < 0.05, vs . model group ( t -test was used to specify differences between two groups at the corresponding time points); (B, E, H) model group at 14 days after transplantation; (C, F, I) transplantation group at 14 days after transplantation. Arrows: NGF protein-, BDNF protein-, and BDNF mRNA-positive cells.

Article Snippet: Primary antibodies were incubated overnight at 4°C in wet box with the following dilutions: rat anti-human BrdU monoclonal antibody (1:150; Beijing Boaosen Biotechnology, China), rabbit anti-mouse BDNF (1:150; Boster, Wuhan, China), NGF (1:150; Boster), VEGF (1:150; Boster), CD34 (1:100; Boster), bFGF (1:100; Beijing Boaosen Biotechnology) polyclonal antibody.

Techniques: Expressing, Transplantation Assay, Immunohistochemistry, Derivative Assay, In Situ Hybridization

Journal: Cell Death & Disease

Article Title: Retrograde transport of neurotrophin receptor TrkB-FL induced by excitotoxicity regulates Golgi stability and is a target for stroke neuroprotection

doi: 10.1038/s41419-025-07990-6

Figure Lengend Snippet: A – D Kinetics of TrkB-FL downregulation. Primary cortical cultures were treated with 100 μM NMDA and its co-agonist 10 μM glycine (hereafter referred to as ‘NMDA’). Immunofluorescence ( A ) and immunoblotting ( B ) used a C-terminal (C-ter) isoform-specific antibody (TrkB-FL Ct) recognizing both the full-length protein (FL) and the intracellular fragment (f32). A Shows TrkB-FL (green) and nuclei (blue, DAPI stain). Arrowheads indicate varicosities in neuronal projections. Scale bar: 20 μm. Insets show cell body details for untreated cells and cells treated with NMDA for 120 min. B Compares the decrease in TrkB-FL and formation of f32 with PSD-95 downregulation, detected using a C-terminal antibody (PSD-95 Ct). Calpain activation was confirmed by the accumulation of characteristic spectrin breakdown products (BDPs; 150 and 145 kDa). Neuron-specific enolase (NSE) served as a loading control for protein normalization. C , D Quantification of normalized TrkB-FL and PSD-95 levels, shown relative to levels in the absence of NMDA (control). Data are represented as means ± SD. Statistical analysis: one-way analysis of variance (ANOVA) followed by Bonferroni post hoc test (** P < 0.01, *** P < 0.001, **** P < 0.0001; 0-90 min, n = 5; 120 min, n = 3). E , F Effect of dynasore preincubation (80 μM, 30 min) on TrkB-FL levels after 2 h NMDA treatment. Data are means ± SD ( n = 4); statistical analysis as above (* P < 0.05). G Sequences of the cell-penetrating neuroprotective peptide (MTFL 457 ) and control peptide (MTMyc). Both contain Tat amino acids (aa) 47–57 (italic), followed by rat TrkB-FL aa 457-471 (light blue) or c-Myc aa 408-421 (dark blue), respectively. H , I Effect of peptide preincubation (25 μM, 30 min) on TrkB-FL shedding and processing during NMDA treatment. Culture media were analyzed using an antibody against the TrkB-FL extracellular domain (panTrkB), which recognizes the ectodomains (ECDs) of all isoforms (TrkB-ECD), to assess receptor shedding by metalloproteinase (MP) activation. Total lysates were analyzed with the TrkB-FL Ct antibody to evaluate TrkB-FL calpain processing via f32 production. Relative TrkB-ECD levels are shown as means ± SD (0-4 h, n = 7; 6 h, n = 4). Statistical analysis: two-way ANOVA followed by Bonferroni post hoc test (** P < 0.01, **** P < 0.0001, comparing each peptide + NMDA vs. peptide alone; # P < 0.05, #### P < 0.0001, comparing MTMyc vs. MTFL 457 at each time point). J , K Effect of NMDA on total and cell-surface pY816-TrkB-FL levels. Cultures were preincubated with peptides as above, then treated briefly with NMDA (1 h) to minimize receptor degradation. Cell-surface proteins were biotin-labeled and precipitated, then compared to corresponding total lysates. Data are represented as means ± SD ( n = 4). Statistical analysis: two-way ANOVA followed by Bonferroni post hoc test ( n.s . = n ot significant; ** P < 0.01, comparing NMDA vs. no NMDA; ## P < 0.01, comparing MTMyc + NMDA vs. MTFL 457 + NMDA).

Article Snippet: Primary antibodies against the following proteins were used: Hrs (Santa Cruz Biotechnology; Cat#sc-271455, RRID:AB_10648901), NSE (Millipore; Cat#AB951, RRID:AB_92390), spectrin alpha chain (Millipore; Cat#MAB1622, RRID:AB_94295), TrkB-FL C-ter (Santa Cruz Biotechnology; sc-11, RRID:AB_632554), TrkB extracellular sequences or panTrkB (Santa Cruz Biotechnology; sc-136990, RRID:AB_2155262), PSD-95 C-ter (Transduction Laboratories; Cat#610496, RRID:AB_2315218), pY816-TrkB-FL (Boster; Cat# P01388 ).

Techniques: Immunofluorescence, Western Blot, Staining, Activation Assay, Control, Labeling

Journal: Clinical and Experimental Immunology

Article Title: The effects of maternal anti-alpha-enolase antibody expression on the brain development in offspring

doi: 10.1093/cei/uxac086

Figure Lengend Snippet: quantitative examination of glial cells in hippocampal and cortical tissues and assessment of cytokine and BDNF levels in the brain tissue of the pups from H-ENO1Ab dams. All the pups were obtained as depicted in . The expression levels of Iba-1 (A) and GFAP (B) in the hippocampal and cortical tissues from the pups of H-ENO1Ab dams and those of the control group on P10 and P40 ( n = 4/group, respectively) were quantitatively assessed by western blot. * P < 0.05 vs. the pups of the control group at the same age. (C) The average levels of cytokines in the supernatants of brain tissue homogenates from the pups of H-ENO1Ab dams and those of the control group on P10 were detected by microarray ( n = 4/group). NS vs. the pups of the control group. (D) The levels of BDNF in the brain tissues of the pups from H-ENO1Ab dams and those of the control group on P10 ( n = 4/group) were detected by western blot. * P < 0.05 vs. the pups of the control group on P10.

Article Snippet: The membranes were incubated at 4°C overnight with the following primary antibodies: rabbit anti-ENO1 antibodies (1:400, BIOSS), rabbit anti-Iba-1 antibodies (1:1000, Abcam), mouse anti-GFAP antibodies (1:1000, CST), rabbit anti-cleaved caspase-3 antibodies (1:1000, CST), rabbit anti-BDNF antibodies (1:500, Wanleibio, China), and rabbit anti-beta-actin antibodies (1:5000, BIOSS).

Techniques: Expressing, Control, Western Blot, Microarray