Journal: bioRxiv

Article Title: Nanoliposomal Omega-3 Fatty Acids Promote Adult Hippocampal Neurogenesis through the BDNF/TrkB Pathway in C57BL/6 Mice

doi: 10.64898/2026.02.24.707750

Figure Lengend Snippet: Protein expression of molecules from the BDNF signaling pathway determined by Western Blot and representative membranes. (A)mBDNF. (B)TrkB. (C)proBDNF. (D)p75 for yogurt treated mice after 4 weeks. (E–F) Representative membranes showing signals for TrkB, mBDNF, pro-BDNF, and p75 in Control and fatty acids-treated mice. Data are expressed as mean ± SD, n =5−6/group. # 0.05 < p < 0.10; *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: Samples (50 μgr in 5× loading buffer) were then loaded into SDS-PAGE gels (12 or 15%) and transferred onto nitrocellulose membranes using the Mini-PROTEAN ® Tetra System (BIO-RAD) for 1 h. Membranes were incubated for 1 h with blocking solution (5% milk in TBST) and then probed overnight at 4°C with mouse anti-BDNF (1:2000; Icosagen; 327-100 clone 3C11), rabbit anti-p75 (1:700; Alomone Labs; ANT-007), rabbit anti-TrkB (1:700; Alomone Labs; ANT-019), and rabbit anti-proBDNF (1:2500; Genocopoeia) in TBST.

Techniques: Expressing, Western Blot, Control

Journal: EMBO Molecular Medicine

Article Title: Impaired migration and lung invasion of human melanoma by a novel small molecule targeting the transmembrane domain of death receptor p75 NTR

doi: 10.1038/s44321-025-00297-1

Figure Lengend Snippet: ( A ) Volcano plot of AraTM screening assay of Screening Set v2010 (8482 compounds) from the Chemical Biology Consortium Sweden ( www.cbcs.se ). Fold change was calculated against vehicle (DMSO) and hits were defined as compounds resulting in greater than ±0.5 fold change in GFP/OD 630 signal without affecting OD 630 by greater than 0.3-fold across three independent runs (red dots). P values were calculated by one-way ANOVA ( N = 3). Compound Div17E5 is indicated. ( B ) Chemical structure of Div17E5. ( C ) Dose response of Div17E5 in the AraTM assay of p75 NTR TMD in comparison with unrelated TMDs from α2β3 integrin and Matrix-2 protein (M2) from the viral envelope of influenza A virus. Results are plotted as mean ± SD ( N = 3). ( D – F ) Comparison of wild-type (WT) p75 NTR TMD and I252A, P253G and V254A mutants in the AraTM assay in response to increasing doses of Div17E5. GFP/OD 630 signal for WT TMD without any drug was set at 100% and all other measurements are relative to that. Results are plotted as mean ± SD ( N = 3). * P = 0.03: ** P = 0.006 (one-way ANOVA followed by Tukey’s multiple comparisons). ( G ) Live cell homo-FRET anisotropy in response to Div17E5 of full-length, wild-type human p75 NTR expressed in COS cells. Shown are representative traces of average anisotropy change after addition of Div17E5 (10 μM) or vehicle. ( H ) Live cell homo-FRET anisotropy in response to Div17E5 of full-length, wild-type human p75 NTR in comparison to I252A p75 NTR TMD mutant. Shown are representative traces of average anisotropy change after addition of Div17E5 (10 μM). .

Article Snippet: For immunoprecipitation, A875 cell extracts were incubated for 16 h at 4 °C on a rotating wheel with 0.8 μg of anti-p75 NTR antibody (ANT-007, Alomone) and then incubated with Sepharose protein-G beads (GE Healthcare).

Techniques: Screening Assay, AraTM Assay, Comparison, Virus, Mutagenesis

Journal: EMBO Molecular Medicine

Article Title: Impaired migration and lung invasion of human melanoma by a novel small molecule targeting the transmembrane domain of death receptor p75 NTR

doi: 10.1038/s44321-025-00297-1

Figure Lengend Snippet: ( A ) Expression of p75 NTR in A875 control (NT, non-targeting) and knock-down (shp75) cells. ( B ) Dose response analysis of cleaved PARP induction by Div17E5 in A875 control (NT) and knock-down (shp75) cells. ( C ) Western blot analysis of cleaved PARP in A875 melanoma cells in response to Div17E5 (10 µM) in the presence or absence of pan-caspase inhibitor Q-VD-pOH (5 µM). Reprobing for GAPDH was used as loading control. The experiment was repeated three times with comparable results. ( D ) Dose-dependent cell viability of A875 control (NT) and knock-down (shp75) cells in response to Div17E5. Results are plotted as mean ± SD ( N = 3). * P = 0.04; ** P = 0.006 (one-way ANOVA followed by Tukey’s multiple comparisons). .

Article Snippet: For immunoprecipitation, A875 cell extracts were incubated for 16 h at 4 °C on a rotating wheel with 0.8 μg of anti-p75 NTR antibody (ANT-007, Alomone) and then incubated with Sepharose protein-G beads (GE Healthcare).

Techniques: Expressing, Control, Knockdown, Western Blot

Journal: EMBO Molecular Medicine

Article Title: Impaired migration and lung invasion of human melanoma by a novel small molecule targeting the transmembrane domain of death receptor p75 NTR

doi: 10.1038/s44321-025-00297-1

Figure Lengend Snippet: ( A ) Chemical structure of Np75-4A22. ( B ) Dose response of Div17E5 and Np75-4A22 in the AraTM assay of p75 NTR TMD. Results are plotted as mean ± SD ( N = 3). ** P = 0.007 (one-way ANOVA followed by Tukey’s multiple comparisons). ( C – E ) Comparison of wild-type p75 NTR TMD and I252A, P253G and V254A mutants in the AraTM assay in response to increasing doses of Np75-4A22. GFP/OD 630 signal without any drug added was set at 100%. Results are plotted as mean ± SD ( N = 3). * P = 0.035; ** P = 0.008 (one-way ANOVA followed by Tukey’s multiple comparisons). ( F – H ) Specific interaction between Np75-4A22 (4 mM) and p75 NTR TMD (1 mM) in bicelles that mimic a lipid bilayer. ( F ) 1 H- 15 N TROSY-HSQC spectra of p75 NTR TMD in DMPC-DH6PC bicelles ( q = 0.5) in the absence (red) and presence (blue) of 4 molar excess Np75-4A22. Cross-peaks undergoing significant chemical shift are indicated with arrows and the corresponding residues labeled. ( G ) The same as ( F ) for the TMD of TrkB receptor tyrosine kinase, showing mostly non-specific interaction. ( H ) Same as ( F ) for the TMD of TNF receptor 2 (TNFR2), showing essentially no interaction. .

Article Snippet: For immunoprecipitation, A875 cell extracts were incubated for 16 h at 4 °C on a rotating wheel with 0.8 μg of anti-p75 NTR antibody (ANT-007, Alomone) and then incubated with Sepharose protein-G beads (GE Healthcare).

Techniques: AraTM Assay, Comparison, Labeling

Journal: EMBO Molecular Medicine

Article Title: Impaired migration and lung invasion of human melanoma by a novel small molecule targeting the transmembrane domain of death receptor p75 NTR

doi: 10.1038/s44321-025-00297-1

Figure Lengend Snippet: ( A ) Representative Western blot analysis (IB) of fascin in p75 NTR immunoprecipitates (IP) and whole cell lysates (WCL) of A875 melanoma cells after overnight treatment with NGF or Np75-4A22 as indicated. The histogram on the right shows quantification expressed as average of three independent experiments, each performed in triplicate ± SD. # P = 0.0091 vs. control (first bar); ** P = 0.0088 vs. NGF alone (second bar) (one-way ANOVA followed by Tukey’s multiple comparisons). ( B ) Representative Western blot analysis of fascin in Triton-insoluble extracts of A875 melanoma cells after overnight treatment with NGF or Np75-4A22 as indicated. The histogram on the right shows quantification expressed as average of three independent experiments, each performed in triplicate ± SD. # P = 0.0412 vs. control (first bar); ** P = 0.0082 vs. NGF alone (second bar) (one-way ANOVA followed by Tukey’s multiple comparisons). ( C ) Photomicrographs of cultured A875 control (NT) and knock-down (shp75) cells treated with NGF or Np75-4A22 as indicated and stained with fluorophor-conjugated phalloidin (green), to reveal actin filaments, and DAPI (blue). White arrows denote some of the filopodia in NGF-treated cells. Scale bar, 5 μM. ( D , E ) Quantification of filopodia per cell in A875 NT ( D ) and A875 shp75 ( E ) cells treated with 100 ng/ml NGF or 1 or 5 μM Np75-4A22 as indicated. Average of three independent experiments, each performed in triplicate, are shown ± SEM. ### P = 0.0004 vs. vehicle (first bar); *** P = 0.0001 vs. NGF alone (second bar). .

Article Snippet: For immunoprecipitation, A875 cell extracts were incubated for 16 h at 4 °C on a rotating wheel with 0.8 μg of anti-p75 NTR antibody (ANT-007, Alomone) and then incubated with Sepharose protein-G beads (GE Healthcare).

Techniques: Western Blot, Control, Cell Culture, Knockdown, Staining

Journal: EMBO Molecular Medicine

Article Title: Impaired migration and lung invasion of human melanoma by a novel small molecule targeting the transmembrane domain of death receptor p75 NTR

doi: 10.1038/s44321-025-00297-1

Figure Lengend Snippet: p75 NTR immunostaining (red) in lung metastasis induced by A875-NT ( A ) or shp75-A875 ( B ) cells counter-stained for human nucleolin (green) and DAPI (white). Scale bar, 100 μM.

Article Snippet: For immunoprecipitation, A875 cell extracts were incubated for 16 h at 4 °C on a rotating wheel with 0.8 μg of anti-p75 NTR antibody (ANT-007, Alomone) and then incubated with Sepharose protein-G beads (GE Healthcare).

Techniques: Immunostaining, Staining

Journal: Biomedicines

Article Title: p75NTR Modulation Reduces Oxidative Stress and the Expression of Pro-Inflammatory Mediators in a Cell Model of Rett Syndrome

doi: 10.3390/biomedicines12112624

Figure Lengend Snippet: Expression of NGF and p75NTR fibroblasts derived from control individuals and Rett patients. ( A ) Total RNA was extracted from healthy control (HC-1) and Rett syndrome (RTT-1) fibroblasts, and the expression levels of ngf were measured by qRT-PCR. n = 3 biological replicates. Data represent means ± SD. ( B ) Immunofluorescence and respective quantitative analysis of NGF in HC and RTT. Cells were fixed in 4% PFA and stained with antibodies against NGF (red). DAPI (blue) was employed for nuclear counterstaining. n = 6 biological replicates. ( C ) qRT-PCR analysis of ngfr ( p75NTR ) in HC-1 and RTT-1 fibroblasts. n = 3 biological replicates. ( D ) Immunofluorescence and respective quantitative analysis of p75NTR immunoreactivity in HC-1 and RTT-1 cells. Cells were fixed in 4% PFA and stained with anti-p75NTR (red). DAPI (blue) was used to counterstain nuclei. n = 5 biological replicates. Data are expressed as mean ± SD. Statistical analysis was performed by using the Student’s unpaired t -test. Statistical significance is indicated as follows: * p < 0.05; ** p < 0.01; *** p < 0.001 vs. DMSO. Images were acquired using the Leica TCS SP8 confocal microscope and Leica Application Suite X (LAS X) software at 40× magnification. Scale bar: 50 µm.

Article Snippet: The anti-NGF and anti-p75NTR antibodies were validated using NGF (BLP-N240, Alomone labs, Jerusalem, Israel) and p75NTR (BLP-NT011 Alomone labs, Jerusalem, Israel) blocking peptides.

Techniques: Expressing, Derivative Assay, Control, Quantitative RT-PCR, Immunofluorescence, Staining, Microscopy, Software

Journal: Biomedicines

Article Title: p75NTR Modulation Reduces Oxidative Stress and the Expression of Pro-Inflammatory Mediators in a Cell Model of Rett Syndrome

doi: 10.3390/biomedicines12112624

Figure Lengend Snippet: Effects of p75NTR modulation on the antioxidant response in RTT fibroblasts. Representative Western blot and densitometric analysis of ( A ) SOD1, ( B ) SOD2, ( C ) Catalase, ( D ) TrxR1, and ( E ) Gpx1 in healthy control fibroblasts (HC-1), Rett syndrome fibroblasts (RTT-1) and RTT fibroblasts treated with 0.1 μM of LM11A-31(RTT-1+LM) for 24 h. n = 3–5 biological replicates. GAPDH was used as a loading control. ( F ) Immunofluorescence and quantification of GSH immunoreactivity in HC-1, RTT-1, and RTT-1+LM experimental groups. Cells were fixed in 4% PFA and stained with antibodies against GSH (red). Nuclear staining was performed with DAPI (blue). n = 12 biological replicates. Data are expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA, followed by Tukey’s post hoc test. Statistical significance is indicated as follows: ** p < 0.01; *** p < 0.001. “a” indicates statistical significance vs. HC-1; “b” indicates statistical significance vs. RTT-1. Images were acquired using the Leica TCS SP8 confocal microscope and Leica Application Suite X (LAS X) software at 40× magnification. Scale bar: 50 µm.

Article Snippet: The anti-NGF and anti-p75NTR antibodies were validated using NGF (BLP-N240, Alomone labs, Jerusalem, Israel) and p75NTR (BLP-NT011 Alomone labs, Jerusalem, Israel) blocking peptides.

Techniques: Western Blot, Control, Immunofluorescence, Staining, Microscopy, Software

Journal: Biomedicines

Article Title: p75NTR Modulation Reduces Oxidative Stress and the Expression of Pro-Inflammatory Mediators in a Cell Model of Rett Syndrome

doi: 10.3390/biomedicines12112624

Figure Lengend Snippet: p75NTR modulation reduces IL-6 and IL-8 expression in RTT fibroblasts. Representative immunofluorescence and respective quantitative analysis of ( A ) IL-6 immunoreactivity in control fibroblasts (HC-1), Rett syndrome fibroblasts (RTT-1), and RTT fibroblasts treated with LM11A-31(RTT-1+LM) at the dose of 0.1 μM for 24 h. Cells were fixed in 4% PFA and stained with antibodies against IL-6 (red). DAPI was used to counterstain nuclei. n = 6 biological replicates. Images were acquired using the Leica TCS SP8 confocal microscope and Leica Application Suite X (LAS X) software at 40× magnification. Scale bar: 50 µm. ( B ) ELISA on IL-6 in culture medium from HC-1, RTT-1, and RTT-1+LM groups treated as abovementioned. n = 4 biological replicates. ( C ) IL-8 immunoreactivity (red) and respective quantitative analysis performed on HC-1, RTT-1, and RTT-1+LM fibroblasts treated as in ( A ). DAPI was used for nuclear staining. n = 6 biological replicates. Scale bar: 50 µm. ( D ) ELISA on secreted IL-8 in conditioned medium from HC-1, RTT-1, and RTT-1+LM cells treated as abovementioned. n = 4 biological replicates. Data are expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA, followed by Tukey’s post hoc test. Statistical significance is indicated as follows: * p < 0.05; ** p < 0.01; *** p < 0.001. “a” indicates statistical significance vs. HC-1; “b” indicates statistical significance vs. RTT-1.

Article Snippet: The anti-NGF and anti-p75NTR antibodies were validated using NGF (BLP-N240, Alomone labs, Jerusalem, Israel) and p75NTR (BLP-NT011 Alomone labs, Jerusalem, Israel) blocking peptides.

Techniques: Expressing, Immunofluorescence, Control, Staining, Microscopy, Software, Enzyme-linked Immunosorbent Assay