recombinant human gfra1  (R&D Systems)

Journal: Frontiers in Pharmacology

Article Title: Enhanced therapeutic potential of paeoniflorin and vitamin B12 in intracerebropeduncle ethidium bromide-induced multiple sclerosis-like pathology

doi: 10.3389/fphar.2026.1792674

Figure Lengend Snippet: The figures illustrate the docking interaction of paeoniflorin with the GDNF receptor (PDB ID: 1AGQ). (A) Shows a surface representation of the GDNF receptor, with the active site highlighted in yellow, where the ligand paeoniflorin (red) is bound. (B) 2D interaction diagram displays key interactions between paeoniflorin and the receptor, with hydrogen bonds and hydrophobic interactions clearly indicated. (C) Provides a 3D view of the docking, showing paeoniflorin (blue) interacting with GDNF, with hydrogen bonds represented by green dashed lines and key regions labeled. (D) Offers a zoomed-in view of the binding site, emphasizing hydrogen bonds and hydrophobic contacts, depicted within a ribbon structure for clarity.

Article Snippet: The ELISA kits for evaluating cellular and molecular targets included GDNF [E-EL-H1495; Elabscience GFRA1 [PKSH033670; Elabscience]; RET [AN00810P; Elabscience], AKT [E-EL-R0807 98T, Elabscience, Wuhan, China] ( ); ERK1/2 [E-AB-70292; Elabscience] ( ); and GSK3-Beta [KLR0989, KRISHGEN, Maharashtra, India] ( ).

Techniques: Labeling, Binding Assay

Journal: Frontiers in Pharmacology

Article Title: Enhanced therapeutic potential of paeoniflorin and vitamin B12 in intracerebropeduncle ethidium bromide-induced multiple sclerosis-like pathology

doi: 10.3389/fphar.2026.1792674

Figure Lengend Snippet: (A–H) PNN neuroprotective role in mitigating EBRO-induced alterations in levels of cellular and molecular targets in MS rat model: GDNF (A) , GFRA1 (B) , AKT (C) , ERK1/2 (D) , GSK3-Beta (E) , in brain homogenates, and GDNF, GFRA1, AKT, ERK1/2, GSK3-Beta in CSF levels (F–H) . Statistical analysis was performed using a one-way ANOVA followed by Tukey’s post hoc test to determine significant differences among groups (A–H) . Data were presented as mean ± standard deviation (SD), with statistical significance set at p < 0.01. Each experimental group consisted of eight wistar rats (n = 8). β v/s Sham Control, Vehicle Control, and PNN Perse; δ v/s EBRO; δα1 v/s EBRO + PNN50; δα2 v/s EBRO + PNN100, EBRO + PNN50; and δα3 v/s EBRO + VB12 (30), EBRO + PNN100, EBRO + PNN50. To identify significant differences between groups, a one-way ANOVA and Tukey’s post hoc test were used for statistical analysis (A–H) . The statistical significance level was set at p < 0.01, and the data were displayed as mean ± standard deviation (SD). There were eight wistar rats (n = 8) in each experimental group. β v/s Sham Control, Vehicle Control, and PNN Perse; δ v/s EBRO; δα1 v/s EBRO + PNN50; δα2 v/s EBRO + PNN100, EBRO + PNN50; and δα3 v/s EBRO + VB12 (30), EBRO + PNN100, EBRO + PNN50.

Article Snippet: The ELISA kits for evaluating cellular and molecular targets included GDNF [E-EL-H1495; Elabscience GFRA1 [PKSH033670; Elabscience]; RET [AN00810P; Elabscience], AKT [E-EL-R0807 98T, Elabscience, Wuhan, China] ( ); ERK1/2 [E-AB-70292; Elabscience] ( ); and GSK3-Beta [KLR0989, KRISHGEN, Maharashtra, India] ( ).

Techniques: Standard Deviation, Control

Journal: Frontiers in Pharmacology

Article Title: Enhanced therapeutic potential of paeoniflorin and vitamin B12 in intracerebropeduncle ethidium bromide-induced multiple sclerosis-like pathology

doi: 10.3389/fphar.2026.1792674

Figure Lengend Snippet: (A–H) PNN neuroprotective role in mitigating EBRO-induced alterations in levels of cellular and molecular targets in MS rat model: GDNF (A) , GFRA1 (B) , AKT (C) , ERK1/2 (D) , GSK3-Beta (E) , in brain homogenates, and GDNF, GFRA1, AKT, ERK1/2, GSK3-Beta in CSF levels (F–H) . Statistical analysis was performed using a one-way ANOVA followed by Tukey’s post hoc test to determine significant differences among groups (A–H) . Data were presented as mean ± standard deviation (SD), with statistical significance set at p < 0.01. Each experimental group consisted of eight wistar rats (n = 8). β v/s Sham Control, Vehicle Control, and PNN Perse; δ v/s EBRO; δα1 v/s EBRO + PNN50; δα2 v/s EBRO + PNN100, EBRO + PNN50; and δα3 v/s EBRO + VB12 (30), EBRO + PNN100, EBRO + PNN50. To identify significant differences between groups, a one-way ANOVA and Tukey’s post hoc test were used for statistical analysis (A–H) . The statistical significance level was set at p < 0.01, and the data were displayed as mean ± standard deviation (SD). There were eight wistar rats (n = 8) in each experimental group. β v/s Sham Control, Vehicle Control, and PNN Perse; δ v/s EBRO; δα1 v/s EBRO + PNN50; δα2 v/s EBRO + PNN100, EBRO + PNN50; and δα3 v/s EBRO + VB12 (30), EBRO + PNN100, EBRO + PNN50.

Article Snippet: The ELISA kits for evaluating cellular and molecular targets included GDNF [E-EL-H1495; Elabscience GFRA1 [PKSH033670; Elabscience]; RET [AN00810P; Elabscience], AKT [E-EL-R0807 98T, Elabscience, Wuhan, China] ( ); ERK1/2 [E-AB-70292; Elabscience] ( ); and GSK3-Beta [KLR0989, KRISHGEN, Maharashtra, India] ( ).

Techniques: Standard Deviation, Control

Journal: bioRxiv

Article Title: Measurement of adhesion and traction of cells at high yield (MATCHY) reveals an energetic ratchet driving nephron condensation

doi: 10.1101/2024.02.07.579368

Figure Lengend Snippet: ( A ) Model schematic of ureteric bud tubule branching in the embryonic kidney. Right, detail of nephrogenic niche anatomy at ureteric bud tips and the role of RET in MAPK activation via ERK. ( B ) Phase contrast micrographs of RET-expressing MDCK cell line as a reductionist model, with and without activation using GDNF and the co-receptor GFRA1. ( C ) Montage of phase and immunofluorescence micrographs and traction force microscopy output (displacement and stress fields) for representative RET-MDCK cells after the indicated treatments on adhesive polyacrylamide substrate. ( D ) Violin plots of traction force distribution for indicated treatments showing means and quartiles (gray lines, n = 8 replicate wells per condition; red points are means of replicates). One-way ANOVA, Tukey’s test, **** p < 0.0001. ( E ) Plots of RET expression and pERK intensity by immunofluorescence upon GDNF/GFRA1 activation vs. untreated control (>1,700 cells combined across n = 8 replicate wells). Traction force is represented by point color and by running average plots (window size of 25 cells).

Article Snippet: To stimulate pERK overexpression, 100 ng/ml of soluble, recombinant human GFRA1 (R&D Systems, AF714-SP) along with 50 ng/ml recombinant human GDNF (R&D Systems, 212-GD-010) were administered for 12 hr.

Techniques: Activation Assay, Expressing, Immunofluorescence, Microscopy, Adhesive, Control

Journal: Cytotherapy

Article Title: Impact of the overexpression of the tyrosine kinase receptor RET in the hematopoietic potential of induced pluripotent stem cells (iPSCs).

doi: 10.1016/j.jcyt.2023.10.003

Figure Lengend Snippet: Figure 1. RET activation with GDNF/GFRa1 has no effect on hematopoietic potential at D13 of iPSCs hematopoietic differentiation. (A) Morphology of non-adherent, round, hematopoietic cells derived from PB33-WT iPSC at D13 of differentiation. (B) Microscope pictures (magnification 10x) of May-Grunwald and Giemsa (MGG) staining at day +13 of floating hematopoietic cells differentiated from PB33-WT. (C) Microscope picture of CFU assay at D+14 of the culture displaying a CFU-GM (white) and BFU-E (red) colony. (DG) Phenotypic analysis of hemato- poietic cells initially gated on live cells. Proportion of CD34 total (D), primitive hematopoietic stem and progenitor cells (CD34+/CD38) (E), HSC phenotype bearing cells (CD34+/CD38/ CD49f+) (F), or total hematopoietic cells (CD45+) (G) at day +13 of hematopoietic differentiation with or without GDNF for 2 different iPSC cell lines (PB33-WT and PB68-WT). (H) CFC assays derived from PB33-WT and PB68-WT with or without GDNF showing the number of colonies per 5000 cells (Means and SD are represented) and the type of colonies (I). All experiments have been performed 5 times. P-values were calculated using a 2-tailed Student’s t-test. ns, not significant. (Color version of figure is available online.)

Article Snippet: For the GDNF/GFRa1 experiment, 100 ng/mL of GDNF & GFRa1 mixed 1:1; (212-GD-010, 714-GR-100; R&D Systems, France) were added to the media.

Techniques: Activation Assay, Derivative Assay, Microscopy, Staining, Colony-forming Unit Assay

Journal: Cancer Research

Article Title: Novel Calcium-Binding Ablating Mutations Induce Constitutive RET Activity and Drive Tumorigenesis

doi: 10.1158/0008-5472.can-22-0834

Figure Lengend Snippet: Figure 2. Three-dimensional fluctuation of CRD caused by CaLM mutation. A, 3D view of CaLM mutations. The cartoon shows an overhead view of the extracellular portion of the RET–GDNF–GFRa1 hexamer (PDB: 6Q2N) with an expanded view of CaLM. The expanded image shows a stick and cartoon representation showing the locations of Ca2þ ions (green sphere) and amino acid residues contributing to Ca2þ ion holding. Mutated residues are highlighted in red. Bottom, CLD, orange; cysteine-rich domain (CRD), gray; glial cell-derived neurotrophic factor (GDNF), light blue; GDNF family receptor alpha1 (GFRa1), pink. B, 3D view of calmodulin mutations. The stick and cartoon model represents the Ca2þ-binding motif of calmodulin (PDB code: 2BE6). Mutated residues in long-QT syndrome patients are highlighted in red. C, Violin plot of binding-free energy between the Ca2þ ion and a RET-CRD monomer at 10 ns intervals. The width of each strip represents the fraction of time points showing a binding-free energy value. The black dashed line represents the median value. Each dotted line represents the interval between the end of the first quartile and the beginning of the fourth quartile. , P < 0.05; , P < 0.0001 by a one-way ANOVA with the Tukey multiple comparisons test. D, Violin plots of RMSD of backbone Ca atoms of CRD monomers. The RMSDs of heavy atoms of each amino acid at 100 ps intervals were calculated with respect to the initial position and plotted. Left, RMSD of WT and native CaLM mutants. Right, artificial alanine mutants. E, Oncogenic properties of artificial CaLM mutants. Left, ERK phosphorylation assay. HEK293H cells transfected with empty vector or WT RET, 110 RET mutants, and kinase-dead mutant (K758M)-expressing plasmids were subjected to immunoblot analysis of pERK, ERK, pRET, RET, and b-actin. Right, NIH3T3 focus formation. The transforming ability of lentiviruses expressing artificial CaLM mutants was examined using parental cells (uninfected), empty virus infection, and lentiviruses to express D567Y (native CaLM mutant), C634R (CCM mutant), and M918T (kinase domain mutant) proteins.

Article Snippet: Recombinant human glial cell-derived neurotrophic factor (GDNF; catalog no. 212-GD) and GFRa1 (catalog no. 714-GR) were purchased from R&D Systems.

Techniques: Mutagenesis, Derivative Assay, Binding Assay, Stripping Membranes, Phospho-proteomics, Transfection, Plasmid Preparation, Expressing, Western Blot, Virus, Infection

Journal: Cancer Research

Article Title: Novel Calcium-Binding Ablating Mutations Induce Constitutive RET Activity and Drive Tumorigenesis

doi: 10.1158/0008-5472.can-22-0834

Figure Lengend Snippet: Figure 3. Illegitimate intermolecular disulfide bond formation induced by CaLM mutations. A, Violin plots showing the interaction energy between the Ca2þ ion and the CRD (left), the RMSD of the CRD (middle), and the SASA of cysteine residues (C565, C570, C581, and C585) in the CRD (right) estimated from MD simulation of the RET– GDNF–GFRa1 extracellular complex. MD simulation was performed in triplicate for each WT and RET mutant, and structures extracted from the 0.5–1 ms trajectories every 20 ps were subjected to calculation. The black dashed line represents the median values. Each dotted line represents the interval between the end of the first quartile and the beginning of the fourth quartile. B, Schematic diagram of the RET protein. CLD, cadherin-like domain; CRD, cysteine-rich domain; KD, kinase domain; green circle, Ca2þ ion-binding sites; dashed line, cell membrane. C, Intermolecular RET dimer formation. Flp-in T-REx 293 cells with or without doxycycline-inducible FLAG-tagged full-length RET cDNAs were cultured for 48 hourswith doxycycline in calcium-containing (left) or calcium-free (right) medium. Whole-cell lysates were prepared under reducing or nonreducing conditions, i.e., presence or absence of dithiothreitol, and resolved by SDS-PAGE, followed by immunoblot analysis using an anti-FLAG antibody. D, Intermolecular dimer formation of CLD4-CRD. GST-tagged RET-CRD-CLD4 polypeptides expressed in Sf21 cells were purified under reducing or nonreducing conditions, i.e., presence or absence of 2-ME, and resolved by SDS-PAGE, followed by immunoblot analysis using an anti-GST antibody. Left, intermolecular dimer formation of WT, C634R, and D567N mutant polypeptides. Right, intermolecular dimer formation of WT RET polypeptides purified with or without calcium chelation, i.e., presence or absence of EDTA. E, RET autophosphorylation and ERK phosphorylation induced by calcium depletion. (Continued on the following page.)

Article Snippet: Recombinant human glial cell-derived neurotrophic factor (GDNF; catalog no. 212-GD) and GFRa1 (catalog no. 714-GR) were purchased from R&D Systems.

Techniques: Mutagenesis, Binding Assay, Membrane, Cell Culture, SDS Page, Western Blot, Phospho-proteomics