Journal: Acta neuropathologica communications

Article Title: BDNF augmentation reverses cranial radiation therapy-induced cognitive decline and neurodegenerative consequences.

doi: 10.1186/s40478-024-01906-9

Figure Lengend Snippet: Fig. 2 Riluzole treatment restores hippocampal BDNF in the irradiated hippocampus. 10–12 weeks old WT male mice received cranial radiation therapy (RT) followed by riluzole (RZ) treatment (13 mg/kg) in drinking water for 6–7 weeks. An ELISA-based quantification of BDNF from the micro-dissected hippocampus showed RT-induced reductions in the RT + Vehicle group. Importantly, RZ treatment in the cranially irradiated mice showed significant restoration of BDNF levels. Data are presented as mean ± SEM (N = 6–10 mice per group). P values were derived from two-way ANOVA and Bonferroni’s multiple comparisons test

Article Snippet: BDNF levels were quantified using a commercially available ELISA kit (E-EL-M0203, Elabscience Biotechnology) and uncoated ELISA plates (Nunc MaxiSorp, Biolegend).

Techniques: Irradiation, Enzyme-linked Immunosorbent Assay, Derivative Assay

Journal: FEBS letters

Article Title: miR-190a inhibits epithelial-mesenchymal transition of hepatoma cells via targeting the long non-coding RNA treRNA.

doi: 10.1016/j.febslet.2015.11.024

Figure Lengend Snippet: Fig. 3. Suppression of treRNA by miR-190a inhibits EMT in HCC. (A) 20 nM miR-190a, 20 nM siRNA duplexes against treRNA (si-treRNA1 or sitreRNA2), 20 nM miR-190a, or NC RNA was transfected into Huh-7 or HepG2 cells. EMT related genes were quantified 72 h post-transfection as indicated. (B) Cells were treated with antagomiR (anti-miR- 190a) or NC RNA as in (A). The mRNA level of different EMT genes was quantified and normalized to b-actin. (C) Cell lysates were blotting with EMT marker E-cadherin (E-cad), Claudin-1 (Claud), Vimentin (Vimen), N-cadherin (N-cad), SNAI1 (Snail). b-Actin was used as endogenous control. The EMT marker expression level in was calculated using Image J and presented as histogram (right panel). (D) Cells were treated with TGF-b for 12 h and transfected with 20 nM miR-190a, 20 nM siRNA duplexes against treRNA (si-treRNA1 or si-treRNA2) or NC RNA subsequently. The morphological change was captured 24 h after transfection.

Article Snippet: Membranes were blocked by 5% BSA or 5% non-fat milk for 3 h at 25 C and incubated with primary antibody against Ago2 (#2897, Cell Signaling, US), Dicer (#5362, Cell Signaling), E-Cadherin (#3195, Cell Signaling), Claudin-1 (#13255, Cell Signaling), Vimentin (#5741, Cell Signaling), N-Cadherin (#13116, Cell Signaling), SNAI1 (#3897, Cell Signaling), or b-actin (C-4, Santa cruz, US) for 12 h at 4 C. Membranes were incubated with secondary antibody diluted in Tris-buffered saline supplemented with 0.5% Tween-20 and 5% BSA or 5% non-fat milk for 1 h at 25 C. The signal was detected with the enhanced chemiluminescence system (Pierce, US) and exposed to X-ray film (Kodak, US).

Techniques: Transfection, Marker, Control, Expressing

Journal: Frontiers in Cellular Neuroscience

Article Title: Hypoxic culture of umbilical cord mesenchymal stem cell-derived sEVs prompts peripheral nerve injury repair

doi: 10.3389/fncel.2022.897224

Figure Lengend Snippet: Cellular localization and uptake of fluorescently labeled sEVs with SCs. (A) Cellular colocalization: SP8 was used to photograph sEVs prelabeled with PKH67 (green fluorescence) with Hoechst 33,342 (blue fluorescence)-stained SC nuclei. (B) Western blot: Detection of ERK1/2, ZEB2, and c-JUN expression levels in NC Schwann cells and after 48-h treatment with hypoxia sEVs. (C) Real-time PCR: Inflammatory and other restoration-related factors (IL-1β, IL-6, TNF-α, etc.) were detected. (D) ELISA test: GDNF NDF and NT-3 in the supernatant of Schwann cell preparation. Statistical significance, * p < 0.05,** p < 0.01, and **** p < 0.0001. *Significant difference.

Article Snippet: The following ELISA kits were: mouse glial cell line-derived neurotrophic factor (GDNF), ELISA kit (CSB-E07341m, CUSABIO), mouse neurotrophin 3 (NT-3), ELISA kit (CSB-E04687m, CUSABIO), and mouse nerve growth factor (NGF), ELISA kit (CSB-E04684m, CUSABIO).

Techniques: Labeling, Fluorescence, Staining, Western Blot, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Molecular Neurobiology

Article Title: Silibinin Alleviates Cognitive Dysfunction, Biochemical Alterations, and Hippocampal Structural Changes Induced by 5 days of Sleep Deprivation with or Without a Subsequent 24-H Recovery in Rats

doi: 10.1007/s12035-026-06195-y

Figure Lengend Snippet: Effect of silibinin administration on A BDNF (ng/ml) and B CaMKII (ng/mg) in sleep-deprived rats with and without consequent 24-h recovery. Data are represented as mean ± SD ( n = 8); group comparisons were performed using one-way ANOVA, followed by Tukey’s multiple-comparisons test. * P < 0.05, ** P < 0.01, and *** P < 0.001 versus the control group; $ P < 0.05, $$ P < 0.01, and $$$ P < 0.001 versus the SD group; # P < 0.05, ## P < 0.01, and ### P < 0.001 versus the SD + R group. SD: sleep-deprived untreated group; SD + R: sleep-deprived untreated group with recovery; SD + Sil: sleep-deprived group treated with silibinin; SD + Sil + R: sleep-deprived group treated with silibinin followed by recovery. BDNF: brain‑derived neurotrophic factor; CaMKII: calcium/calmodulin‑dependent protein kinase II

Article Snippet: Furthermore, hippocampal tissue homogenates were prepared to determine BDNF (Boster Biological Technology, Pleasanton, CA, USA; Cat. No. EK0308) and CaMKII (BlueGene Biotech, Shanghai, China; Cat. No. E02C0682) using commercially available ELISA kits supplied by Bio-KITS (Cairo, Egypt), according to the manufacturers' instructions.

Techniques: Control

Journal: bioRxiv

Article Title: Cholesterol recognition motifs in the transmembrane domain of the tyrosine kinase receptor family: the case for TRKB

doi: 10.1101/734012

Figure Lengend Snippet: TRKB and cholesterol interaction. (a,b) Snapshots of the interaction between TRKB and cholesterol at the TMR (see supplement video) indicating that cholesterol interacts with the OH- group (red) in Y433 (blue) in TRKB.TMR (green). (c) The interaction between the TRKB CARC motif and cholesterol happens preferentially at the C-alpha in Y433 residues, as indicated by coarse-grained molecular dynamics simulations. (d) The binding of biotinylated BDNF (bBDNF) and TRKB is modulated by cholesterol in a bell-shaped fashion. Red rectangle area is expanded in (e) for the comparison between BDNF 1ng/ml (black circles) with BDNF 0 (ctrl, open circles). Data expressed as Mean/SEM of (c) cholesterol occupancy; or from binding normalized by (d) chol 0/BDNF10ng/ml or (e) chol 0/BDNF 1ng/ml. ∗p<0.05 from control (BDNF 0 or ctrl, C427).

Article Snippet: The plates were washed 3x with PBS buffer, and a mixture of biotinylated BDNF (bBDNF, 0, 0.1, 1 or 10ng/ml, Alomone Labs, #B-250-B) and cholesterol (0, 20, 50 or 100μM) was added for 1h at room temperature, followed by washing with PBS.

Techniques: Binding Assay, Comparison, Control