Journal: Journal of Toxicology

Article Title: Conditioned Medium Reconditions Hippocampal Neurons against Kainic Acid Induced Excitotoxicity: An In Vitro Study

doi: 10.1155/2014/194967

Figure Lengend Snippet: HEK-CM stimulates host's cell survival factors BDNF and Bcl-2. (a) Photomicrograph represents the expression of BDNF in different treatment groups. Scale Bar 200 μ m. (b) Bar diagram signifies the fold changes in BDNF fluorescence intensity as compared to NC. (c) MTT Assay. Bar diagram represents the viable cells in different treatment groups. Neutralization of BDNF in HEK-CM diminishes the neuroprotective potential of HEK-CM against excitotoxicity. Nevertheless, significant neuroprotection was observed in KA + CM with anti-BDNF group as compared to KA group suggesting that other growth factors/cytokines in CM might be contributing to the observed neuroprotection. Normal control (NC), normal control exposed to HEK conditioned medium (CM), Kainic acid alone treated (KA), Kainic acid and HEK conditioned medium cotreated (KA + CM), and Kainic acid and BDNF neutralized HEK conditioned medium cotreated (KA + CM with anti-BDNF). Data represented as mean ± SEM. * indicates comparison between NC and treatment; + indicates comparison between KA lesion and treatment; * P < 0.05; *** P < 0.001; +++ P < 0.001.

Article Snippet: Following several washes in PBS, cells were incubated in blocking buffer (3% bovine serum albumin in PBS) for 30 minutes at room temperature and were incubated with primary monoclonal antibodies against brain derived neurotrophic factors (BDNF) (1 : 200; Developmental studies hybridoma bank, Iowa, USA) overnight at 4°C.

Techniques: Expressing, Fluorescence, MTT Assay, Neutralization, Control, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Effects of Strontium-Doped β-Tricalcium Scaffold on Longitudinal Nuclear Factor-Kappa Beta and Vascular Endothelial Growth Factor Receptor-2 Promoter Activities during Healing in a Murine Critical-Size Bone Defect Model

doi: 10.3390/ijms21093208

Figure Lengend Snippet: Relative protein expression of Osterix in the scaffold. ( a ) The images represent sections that have been immunohistochemically stained against Osterix (Osx) in the vicinity of newly formed tissue in the scaffold (red square) in both the β-TCP (left) and β-TCP + Sr (right) groups. Green arrows indicate Osx-positive cells shown up by red staining. ( b ) The ratio of Osx-positive to total cells in the β-TCP group was significantly higher than in the β-TCP + Sr group (n ≥ 3; p value is given as p ≤ 0.05. Asterisks indicate statistical differences within the groups).

Article Snippet: Afterwards, sections were incubated for 24 h at 4 °C with primary antibodies against Osterix (Osx, Santa Cruz Biotechnology, Heidelberg, Germany; Cat# Sc22536-R; 1:250 in 1.5% BSA in TBS).

Techniques: Expressing, Staining

Journal: Molecular medicine reports

Article Title: Effect of exercise-induced neurogenesis on cognitive function deficit in a rat model of vascular dementia.

doi: 10.3892/mmr.2016.4891

Figure Lengend Snippet: Figure 6. Treadmill exercise promotes mature BDNF expression. (A) Representative photomicrographs of western blot analysis showing BDNF levels in the hippocampus of rats in the SC, SC+EX, 2VO and 2VO+EX groups. (B) The intensity of each band was densitometrically determined and normalized against β‑actin. Results are presented as the mean ± standard error of the mean, n=4. **P<0.01, ***P<0.001 vs. SC; #P<0.05 vs. 2VO. SC, sham‑surgery group; SC+EX, sham‑surgery group subjected to treadmill exercise (15 m/min, 30 min/day for 4 weeks); 2VO, 2VO surgery group; 2VO+EX, 2VO‑surgery group subjected to treadmill exercise (15 m/min, 30 min/day for 4 weeks); BDNF, brain‑derived neurotrophic factor.

Article Snippet: They were then incubated overnight at 4 ̊C with the following primary antibodies: Rabbit polyclonal anti-rat BDNF antibody (cat. no. sc-546; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA; 1:500), rabbit polyclonal anti-rat phosphorylated CREB antibody (cat. no. 9191; Cell Signaling Technology, Inc., Danvers, MA, USA; 1:1,000), rabbit polyclonal anti-rat-CREB antibody (cat. no. 9197; Cell Signaling Technology Inc.; 1:1,000), mouse monoclonal anti-rat β-actin antibody (cat. no. A5441; Sigma-Aldrich; 1:5,000).

Techniques: Expressing, Western Blot

Journal: Cell Death Discovery

Article Title: PGC-1a mediated mitochondrial biogenesis promotes recovery and survival of neuronal cells from cellular degeneration

doi: 10.1038/s41420-024-01953-0

Figure Lengend Snippet: A Representative bright field (BF) and immunofluorescence images of 5% EtOH treatment (vol/vol, 1 and 3 h) induced neurite retraction, and its regeneration after washing EtOH (Washed, 4 and 20 h) in neuronal PC12 cells. Green: neuron-specific β-III tubulin; Blue: DAPI. Scale bar: 50 µm. B , C Quantification results of mean neurite length per cell and the percentage of cells with neurites. *** P < 0.001, vs. control group; ## P < 0.01, ### P < 0.001, vs. EtOH 3 h group. EtOH ethanol.

Article Snippet: For immunostaining, cells were incubated with primary antibodies against neuron-specific β-III tubulin (MAB1195, R&D systems, USA), LC3B (NB100-2220, Novus Biologicals, USA), TOM20 (42406S, Cell Signaling Technology), and Drp1 (8570S, Cell Signaling Technology) overnight at 4 °C.

Techniques: Immunofluorescence, Control

Journal: iScience

Article Title: Investigating microglia-neuron crosstalk by characterizing microglial contamination in human and mouse patch-seq datasets

doi: 10.1016/j.isci.2023.107329

Figure Lengend Snippet:

Article Snippet: Patch-Seq of neurons in mouse primary visual cortex , Gouwens et al., , Allen Institute for Brain Science Cell Types Database.

Techniques: Software

Journal: International Journal of Molecular Sciences

Article Title: Differences in Functional Expression of Connexin43 and Na V 1.5 by Pan- and Class-Selective Histone Deacetylase Inhibition in Heart

doi: 10.3390/ijms19082288

Figure Lengend Snippet: Changes in cardiac Cx43 and Na V 1.5 protein levels by panobinostat, entinostat and ricolinostat. ( A ) Representative Western blot of lysed NMVMs treated with 0, 25, 100, 500, and 2500 nM panobinostat for 24 h. Ac-tubulin is acetylated α-tubulin, Ac-H3 is acetylated histone 3, and α-tubulin was used as a loading control. ( B ) Densitometry scans of Cx43 Western blots ( n = 3) to quantify the reduction in Cx43 protein levels with increasing concentrations of panobinostat (* p < 0.05). ( C ) Densitometry scans of Na V 1.5 Western blots ( n = 3) quantifying the statistically significant decrease in Na V 1.5 protein levels with higher concentrations of panobinostat (* p < 0.05). ( D ) A representative Western blot of lysed ventricular myocytes treated with 1 μM entinostat (MS-275) or 25 nM ricolinostat for 24 h. Note that MS-275 only increased the Ac-H3 signal while ACY-1215 only increased the Ac-α-tubulin signal, consistent with their class I and HDAC6 inhibitory activities. ( E ) Densitometry scans of 1 μM entinostat Western blots ( n = 4) illustrating no significant changes in Cx43 or Na V 1.5 protein levels with entinostat, a class I HDAC-selective inhibitor. ( F ) Densitometry scans of 25 nM ricolinostat Western blots ( n = 3) illustrating no significant changes in Cx43 or Na V 1.5 protein levels with ricolinostat.

Article Snippet: Primary antibodies used in this study include rabbit anti-Cx43 (AB1728, Merck Millipore, Billerica, MA, USA), mouse anti-Cx43 (AB1727, Merck Millipore), mouse anti-α-tubulin (# T5168, Sigma-Aldrich, St. Louis, MO, USA), rabbit anti-acetylated H3 (# 06-599, Merck Millipore), rabbit anti-acetylated-α-tubulin (BML-SA452-0100, Enzo Life Sciences, Farmingdale, NY, USA) and rabbit anti-Na V 1.5 (# ACC-001, Alomone Labs, Jerusalem, Israel).

Techniques: Western Blot