ngf prongf neutralizing antibody  (Alomone Labs)

Journal: Cells

Article Title: ProNGF/p75NTR Axis Drives Fiber Type Specification by Inducing the Fast-Glycolytic Phenotype in Mouse Skeletal Muscle Cells

doi: 10.3390/cells9102232

Figure Lengend Snippet: Effect of proNGF administration and anti-NGF/proNGF addition on SCDM differentiation. ( A ) SCDMs were allowed to differentiate into myotubes in the differentiation medium (DM), in presence or not of proNGF (100 ng/mL) for 72 h. Cells were observed with bright field microscopy and three images for every single experiment were captured. ( B ) SCDMs were treated with anti-NGF/proNGF (500 ng/mL) and induced to differentiate in DM for 72 h. Three images for each experimental group were captured under a brightfield microscope, and then analyzed to evaluate myotube length, myotube diameter and myotube number. ( C ) SCDMs were induced to differentiate in the presence of proNGF (100 ng/mL) or anti-NGF/proNGF (500 ng/mL) for 72 h. Once differentiation was completed, the cells were fixed in paraformaldehyde (PFA) 4% and incubated overnight with anti-MyHC (green) to evaluate the fusion index by immunofluorescence. DAPI (blue) was used to counterstain the nuclei. Fusion index was calculated as the percentage of nuclei incorporated into myotubes on the total nuclei present in each field. All the experiments consisted of at least three biological replicates. Scale bars: 100 μm. Values are expressed as the mean ± SD. Statistical analysis was performed by using the Student’s t test. * p

Article Snippet: Complete differentiation was achieved after 72 h. The beginning and duration of the treatment with 500 ng/mL of NGF/proNGF neutralizing antibody (Alomone labs, Jerusalem, Israel; ALM-006), 100 ng/mL of recombinant mouse proNGF (Alomone labs, Jerusalem, Israel; N-250), and/or 100 nM of LM11A-31 (Sigma Aldrich, Milan, Italy; SML0664) is indicated in the figure legends.

Techniques: Microscopy, Incubation, Immunofluorescence

Journal: Cells

Article Title: ProNGF/p75NTR Axis Drives Fiber Type Specification by Inducing the Fast-Glycolytic Phenotype in Mouse Skeletal Muscle Cells

doi: 10.3390/cells9102232

Figure Lengend Snippet: Effect of proNGF administration and anti-NGF/proNGF addition on SCDM proliferation. ( A ) SCDMs were cultured in a proliferation medium and treated with proNGF (100 ng/mL) up to 48 h. Cells were counted at both 24 and 48 h to evaluate cell growth. ( B ) SCDM cell growth was evaluated as in A, in both control cells and SCDMs treated with neutralizing anti-NGF/proNGF antibody (500 ng/mL) for 48 h. ( C ) EdU staining and the ratio of EdU + cells from proliferating SCDMs, kept in a proliferation medium in the presence or not of proNGF (100 ng/mL) or anti-NGF/proNGF (500 ng/mL) for 48 h. DAPI was used to counterstain the nuclei. Three images for each experimental group were captured under a fluorescence microscope, and then analyzed to evaluate the percentage of EdU + cells. All the experiments consist of at least three biological replicates. Scale bar: 50 μm. Values are expressed as the mean ± SD. For cell proliferation, statistical analysis was performed by using two-way ANOVA, whereas the ratio of EdU + cells was analyzed by one-way ANOVA followed by Tukey’s post hoc test.

Article Snippet: Complete differentiation was achieved after 72 h. The beginning and duration of the treatment with 500 ng/mL of NGF/proNGF neutralizing antibody (Alomone labs, Jerusalem, Israel; ALM-006), 100 ng/mL of recombinant mouse proNGF (Alomone labs, Jerusalem, Israel; N-250), and/or 100 nM of LM11A-31 (Sigma Aldrich, Milan, Italy; SML0664) is indicated in the figure legends.

Techniques: Cell Culture, Staining, Fluorescence, Microscopy

Journal: Biomedicines

Article Title: Development and Biological Characterization of a Novel Selective TrkA Agonist with Neuroprotective Properties against Amyloid Toxicity

doi: 10.3390/biomedicines10030614

Figure Lengend Snippet: ENT-A013, ENT-A013E and ENT-A013Z protect PC12 cells against serum deprivation-induced cell death in a TrkA-dependent manner. PC12 cells were starved of serum and treated with NGF, ENT-A013, ENT-A013E, ENT-A013Z or vehicle in the presence or absence of a specific TrkA inhibitor GW441756 for 24 h and CellTox assay was used to quantify cell death. CellTox positive cell number was normalized to total Hoescht positive cell number in each image. Representative images from 4–9 independent experiments are shown ( A ) and quantification ( B ). Data represent ±SEM. One-way ANOVA, Tukey’s test correction: ns: non-significant; * and # p

Article Snippet: For neurite outgrowth assay, cells were cultured for 5 days in vitro with medium deprived of NGF in the presence of ENT-A013 (500 nM, every 48 h), along with NGF-neutralizing antibody.

Techniques:

Journal: Biomedicines

Article Title: Development and Biological Characterization of a Novel Selective TrkA Agonist with Neuroprotective Properties against Amyloid Toxicity

doi: 10.3390/biomedicines10030614

Figure Lengend Snippet: Docking poses of the two ENT-A013 isomers, ENT-A013E and ENT-A013Z, in sites 1a and 1b at the two symmetry-related TrkA-D5—NGF interfaces of the neurotrophin-receptor complex. The close-up views show residues lining the two sites. TrkA-D5 and NGF are shown in green and blue cartoon representation, respectively, with selected residues in stick representation colored by atom type. The compounds are shown in orange stick representation.

Article Snippet: For neurite outgrowth assay, cells were cultured for 5 days in vitro with medium deprived of NGF in the presence of ENT-A013 (500 nM, every 48 h), along with NGF-neutralizing antibody.

Techniques: