gdnf family receptorα1 gfrα1  (R&D Systems)

Journal: The World Journal of Men's Health

Article Title: Bisphenol Analogs Downregulate the Self-Renewal Potential of Spermatogonial Stem Cells

doi: 10.5534/wjmh.230166

Figure Lengend Snippet: BPA, BPF, and BPS do not alter specific biomarker expression. Fluorescence microscopic images of immunocytochemistry analysis of BPA-, BPF-, and BPS-treated (0, 25, and 100 µM) spermatogonial stem cells (SSCs) (A), scale bar=50 µm, 40X magnification. Localization of specific proteins GFRα1 and PLZF (undifferentiated spermatogonia, red), VASA (germ cells, red), and c-Kit (differentiated spermatogonia, red), and DAPI (nuclei, blue), GFP (cultured germ cells enriched for SSCs, green). The ratio of marker-expressing cells per GFP + germ cells is presented by bar graph as mean±standard error of the mean (SEM) from three independent experiments. (B) Graphical representation of A is shown as the ratio of marker-expressing cells to GFP + germ cells presented as mean±SEM from three independent experiments. (C) Relative gene expression of undifferentiation biomarker genes ( Id4 , Bcl6b , Lhx1 , and Bmi1 ) are presented by bar graph as mean fold change±SEM from four independent experiments. Statistical analysis with one-way ANOVA and multiple comparisons by Dunnett’s test was conducted. There were no significant differences between treatments. BPA: bisphenol-A, BPF: bisphenol-F, BPS: bisphenol-S.

Article Snippet: SSCs were maintained in mouse serum-free medium (mSFM) supplemented with glial cell line-derived neurotrophic factor (GDNF) (10 ng/mL, 212-GD-50; R&D Systems), GDNF family receptorα1 (GFRα1) (75 ng/mL, 560-GR-100; R&D Systems), and basic fibroblast growth factor (1 ng/mL, 354060; BD Biosciences), as previously reported [ ].

Techniques: Biomarker Assay, Expressing, Fluorescence, Immunocytochemistry, Cell Culture, Marker

Journal: International Journal of Molecular Sciences

Article Title: Glial-Cell-Line-Derived Neurotrophic Factor Promotes Glioblastoma Cell Migration and Invasion via the SMAD2/3-SERPINE1-Signaling Axis

doi: 10.3390/ijms251810229

Figure Lengend Snippet: SERPINE1 is increased in GBMs. ( A ) GO enrichment analysis of upregulated DEGs in C6 cells treated with GDNF. The top 30 significant GO terms were exhibited according to p -value. ( B ) SERPINE1 mRNA expressions were compared between the GBM and normal tissues from the UALCAN database. ( C ) The overall survival was analyzed to evaluate the association between the mRNA level of SERPINE1 and the outcome of GBM patients by the Oncomine website. ( D , E ) The protein expressions of SERPINE1 in GBM tissues and cells were determined by Western blotting. NB: normal brain; RAs: rat astrocytes; HAs: human astrocytes (vs. NB: **, p < 0.01. vs. RAs: *, p < 0.05. vs. HAs: #, p < 0.05; ##, p < 0.01).

Article Snippet: The GDNF recombinant protein (human: P39905-1, MedChemExpress, Monmouth Junction, NJ, USA; rat: QP5504, enQuire Bio, Denver, CO, USA) was dissolved with PBS to a concentration of 10 μg/mL, and stored at −80 °C.

Techniques: Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Glial-Cell-Line-Derived Neurotrophic Factor Promotes Glioblastoma Cell Migration and Invasion via the SMAD2/3-SERPINE1-Signaling Axis

doi: 10.3390/ijms251810229

Figure Lengend Snippet: GDNF promotes SERPINE1 expressions and secretion in GBM cells. ( A ) Western blotting (left) was used to detect the protein expressions of SERPINE1 in C6 and U251 cells treated with various doses of GDNF (0, 20, 40, 80, 100 ng/mL) for 48 h. The statistical analysis (right) of the left bands was performed. ( B ) The contents of SERPINE1 were tested in C6 and U251 cells after 24 and 48 h of treatment with different concentrations of GDNF using ELISA kits (vs. 0 ng/mL: *, p < 0.05; **, p < 0.01).

Article Snippet: The GDNF recombinant protein (human: P39905-1, MedChemExpress, Monmouth Junction, NJ, USA; rat: QP5504, enQuire Bio, Denver, CO, USA) was dissolved with PBS to a concentration of 10 μg/mL, and stored at −80 °C.

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Glial-Cell-Line-Derived Neurotrophic Factor Promotes Glioblastoma Cell Migration and Invasion via the SMAD2/3-SERPINE1-Signaling Axis

doi: 10.3390/ijms251810229

Figure Lengend Snippet: SERPINE1 knockdown strikingly suppresses the migration and invasion of GBM cells enhanced by GDNF. ( A , B ) qPCR and Western blotting were applied to assess the knockdown efficiency of SERPINE1 in C6 and U251 cells. ( C , D ) Wound healing assay was performed to examine the effects of SERPINE1 deficiency on cell migration in C6 and U251 cells separately treated with 80 and 20 ng/mL GDNF for 48 h. Bar = 100 µm. ( E , F ) The migratory abilities were detected by transwell migration assay in SERPINE1 silenced C6 and U251 cells treated with 80 and 20 ng/mL GDNF for 48 h, respectively. Bar = 50 µm. ( G ) Transwell invasion assay was conducted to evaluate the influences of SERPINE1 knockdown on cell invasion in C6 and U251 cells after 48 h of separate treatment with 80 and 20 ng/mL GDNF. Bar = 50 µm (*, p < 0.05; **, p < 0.01; ***, p < 0.001).

Article Snippet: The GDNF recombinant protein (human: P39905-1, MedChemExpress, Monmouth Junction, NJ, USA; rat: QP5504, enQuire Bio, Denver, CO, USA) was dissolved with PBS to a concentration of 10 μg/mL, and stored at −80 °C.

Techniques: Knockdown, Migration, Western Blot, Wound Healing Assay, Transwell Migration Assay, Transwell Invasion Assay

Journal: International Journal of Molecular Sciences

Article Title: Glial-Cell-Line-Derived Neurotrophic Factor Promotes Glioblastoma Cell Migration and Invasion via the SMAD2/3-SERPINE1-Signaling Axis

doi: 10.3390/ijms251810229

Figure Lengend Snippet: GDNF upregulates SERPINE1 via SMAD2/3 phosphorylation in GBM cells. ( A , B ) The levels of SMAD2/3 phosphorylation were analyzed in C6 and U251 cells separately treated with 80 and 20 ng/mL GDNF for 0, 1, 2, and 4 h using Western blotting. ( C ) The effects of SMAD2/3 deficiency on SERPINE1 protein expressions were assessed in C6 cells in the presence of 80 ng/mL GDNF for 48 h by Western blotting (*, p < 0.05; **, p < 0.01; ***, p < 0.001).

Article Snippet: The GDNF recombinant protein (human: P39905-1, MedChemExpress, Monmouth Junction, NJ, USA; rat: QP5504, enQuire Bio, Denver, CO, USA) was dissolved with PBS to a concentration of 10 μg/mL, and stored at −80 °C.

Techniques: Phospho-proteomics, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Glial-Cell-Line-Derived Neurotrophic Factor Promotes Glioblastoma Cell Migration and Invasion via the SMAD2/3-SERPINE1-Signaling Axis

doi: 10.3390/ijms251810229

Figure Lengend Snippet: GDNF accelerates GBM growth in vivo. ( A , B ) The tumor tissues were exfoliated and weighed at 14 days after subcutaneous tumor formation. ( C ) The mean tumor volumes were calculated every 2 days until 14 days. ( D ) Hematoxylin-Eosin (HE) staining of the tumor tissues was performed (×20). ( E , F ) IHC staining was used to detect the levels of Ki-67, GFAP, MMP2 and MMP9 (×20). n = 5 (*, p < 0.05; **, p < 0.01. *** p < 0.001).

Article Snippet: The GDNF recombinant protein (human: P39905-1, MedChemExpress, Monmouth Junction, NJ, USA; rat: QP5504, enQuire Bio, Denver, CO, USA) was dissolved with PBS to a concentration of 10 μg/mL, and stored at −80 °C.

Techniques: In Vivo, Staining, Immunohistochemistry

Journal: iScience

Article Title: The regulation of enteric neuron connectivity by semaphorin 5A is affected by the autism-associated S956G missense mutation

doi: 10.1016/j.isci.2024.109638

Figure Lengend Snippet:

Article Snippet: Recombinant Rat GDNF, CF , R&D Systems - bio-techne , Cat: 512-GF-010/CF.

Techniques: Recombinant, Western Blot, Mutagenesis, Bicinchoninic Acid Protein Assay, Software, Membrane