human gfra1 r d systems  (R&D Systems)

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: Cell reports

Article Title: Dissecting the spermatogonial stem cell niche using spatial transcriptomics

doi: 10.1016/j.celrep.2023.112737

Figure Lengend Snippet: (A) Spatial mapping of major testicular cell types (left) and transcriptional states of spermatogonia (right) in a human testis sample. ES, elongating/elongated spermatid; RS, round spermatid; SPC, spermatocyte; SPG, spermatogonium. Scale bar, 300 μm. (B) The spatial relationship between the five SPG transcriptional subtypes as quantified by the neighborhood enrichment score in two independent human Slide-seq datasets. (C) Differentially expressed LR pairs among SPG1-neighboring cell type pairs. (D) Spatial expression patterns of selective LR pairs enriched in myoid cell-SPG1 (upper panel) and endothelial cell-SPG1 (lower panel) pairs. Scale bar, 320 μm. (E) Protein expression of VWF and EFNA1 in a human testicular sample. The white dashed line outlines the blood vessel. Yellow arrowheads denote protein co-localization. Scale bar, 8 μm. (F) Bar graph showing the number of GFRA1 + ; KIT ‒ human SPG on day 7 normalized against that from day 0. Cells were treated with recombinant human ENFA protein at 0, 3, and 6 ng/mL, respectively. Data from 4 patient samples are shown.

Article Snippet: Cells were washed and incubated in 1X DPBS (DPBS-S, 14200-075, Gibco) containing 1% FBS (10082-147, Gibco), goat anti-GFRa1 antibody (5μL/10 6 cells; AF714, R&D Systems) and APC mouse anti-human CD117/KIT antibody (5μL/10 6 cells, 550412, BD Biosciences) for 25 min on ice.

Techniques: Expressing, Recombinant

Journal: Cell reports

Article Title: Dissecting the spermatogonial stem cell niche using spatial transcriptomics

doi: 10.1016/j.celrep.2023.112737

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Cells were washed and incubated in 1X DPBS (DPBS-S, 14200-075, Gibco) containing 1% FBS (10082-147, Gibco), goat anti-GFRa1 antibody (5μL/10 6 cells; AF714, R&D Systems) and APC mouse anti-human CD117/KIT antibody (5μL/10 6 cells, 550412, BD Biosciences) for 25 min on ice.

Techniques: Recombinant, DNA Library Preparation, Software