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: Antibody against human p75 NTR extracellular domain used for immunoprecipitation was obtained from Alomone Labs (#ANT-007); the one used for immunofluorescence was from Promega (#G323A).

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

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: Antibody against human p75 NTR extracellular domain used for immunoprecipitation was obtained from Alomone Labs (#ANT-007); the one used for immunofluorescence was from Promega (#G323A).

Techniques: Expressing, Control, Knockdown, Analysis, 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 ) Phase contrast photomicrographs of wound healing cell motility assay in A875 cells treated with vehicle or Div17E5 in the presence or absence of pan-caspase inhibitor Q-VD-pOH. Size bar (bottom right), 2 mm. ( B , C ) Time course of wound healing in A875-NT control ( B ) and knock-down shp75 ( C ) cells in response to Div17E5 in the presence or absence of Q-VD-pOH. Results are plotted as mean ± SD ( N = 3). * P = 0.044; ** P = 0.008 (one-way ANOVA followed by Tukey’s multiple comparisons). ( D ) Boyden chamber analysis of A875 cell chemotaxis in response to NGF in the presence or absence or Div17E5 or pan-caspase inhibitor Q-VD-pOH. Results are plotted as mean ± SD ( N = 3). ** P = 0.004; *** P = 0.00076 (one-way ANOVA followed by Tukey’s multiple comparisons). ( E ) Boyden chamber analysis of cell chemotaxis of knock-down (shp75) A875 cells in response to NGF or serum (FBS). Results are plotted as mean ± SD ( N = 3). ** P = 0.008 (one-way ANOVA followed by Tukey’s multiple comparisons). .

Article Snippet: Antibody against human p75 NTR extracellular domain used for immunoprecipitation was obtained from Alomone Labs (#ANT-007); the one used for immunofluorescence was from Promega (#G323A).

Techniques: Phase Contrast, Motility Assay, Control, Knockdown, Analysis, Chemotaxis Assay, Wound Healing Assay

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: Antibody against human p75 NTR extracellular domain used for immunoprecipitation was obtained from Alomone Labs (#ANT-007); the one used for immunofluorescence was from Promega (#G323A).

Techniques: AraTM Assay, Comparison, Labeling, Nuclear Magnetic Resonance

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 ) Western blot analysis of cleaved PARP in A875 melanoma cells in response to increasing concentrations (0.5 to 5 µM) of Div17E5 and Np75-4A22. A potent inducer of apoptosis, anisomycin (1 μM), was used as positive control. Reprobing for GAPDH was used as loading control. The histogram on the right shows quantification expressed as average of three independent experiments, each performed in triplicate ± SEM. ( B ) Time course of wound healing in A875 control (NT) and knock-down (shp75) cells in response to low micromolar and submicromolar concentrations of Div17E5 (left side) or Np75-4A22 (right side). Results are plotted as mean ± SEM ( N = 3). * P = 0.035 (one-way ANOVA followed by Tukey’s multiple comparisons). ( C ) Boyden chamber analysis of A875 cell chemotaxis in response to NGF (50 ng/ml) in the presence or absence or Div17E5 (left side graph) or Np75-4A22 (right side graph). Results are plotted as mean ± SEM ( N = 3 independent experiments each performed in triplicate). # P = 0.0465 vs. no NGF; * P = 0.0435 vs. no drug (one-way ANOVA followed by Tukey’s multiple comparisons). ( D ) Boyden chamber analysis of A875 cell chemotaxis in response to NGF (50 ng/ml) in the presence of submicromolar concentrations of Np75-4A22. Results are plotted as mean ± SEM ( N = 2 independent experiments each performed in triplicate). # P = 0.0477 vs. no NGF; * P = 0.0462 and ** P = 0.0078 vs. NGF without drug (one-way ANOVA followed by Tukey’s multiple comparisons). ( E ) Boyden chamber analysis of WM115 cell chemotaxis in response to NGF (100 ng/ml) in the presence of Np75-4A22 (1 μM). Results are plotted as mean ± SEM ( N = 3 independent experiments each performed in triplicate). ### P = 0.0004 vs. vehicle; *** P = 0.0004 vs. NGF without drug (one-way ANOVA followed by Tukey’s multiple comparisons). .

Article Snippet: Antibody against human p75 NTR extracellular domain used for immunoprecipitation was obtained from Alomone Labs (#ANT-007); the one used for immunofluorescence was from Promega (#G323A).

Techniques: Western Blot, Analysis, Positive Control, Control, Knockdown, Chemotaxis Assay

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: Antibody against human p75 NTR extracellular domain used for immunoprecipitation was obtained from Alomone Labs (#ANT-007); the one used for immunofluorescence was from Promega (#G323A).

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

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: Antibody against human p75 NTR extracellular domain used for immunoprecipitation was obtained from Alomone Labs (#ANT-007); the one used for immunofluorescence was from Promega (#G323A).

Techniques: Immunostaining, 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: Reagents and tools table

Article Snippet: Antibody against human p75 NTR extracellular domain used for immunoprecipitation was obtained from Alomone Labs (#ANT-007); the one used for immunofluorescence was from Promega (#G323A).

Techniques: Bacteria, Mouse Assay, Recombinant, Software, Analysis

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: 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: Reagents and tools table

Article Snippet: P75ntr ECD , Alomone Labs , ANT-007.

Techniques: