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

Journal: Molecular Biology of the Cell

Article Title: Interplay of septin amphipathic helices in sensing membrane-curvature and filament bundling

doi: 10.1091/mbc.e20-05-0303

Figure Lengend Snippet: FIGURE 3: Genetic analyses of Shs1 and Cdc12 AH domains. (A) Viability of cdc12-6 cells expressing indicated SHS1 alleles expressed from the endogenous locus (with 3xHA tag, unless indicated with GFP tag, which lacks 3xHA) based on tetrad dissections at permissive temperature (24°C). Cells from genotypes labeled in blue appeared normal, without obvious septin defects. Genotypes in orange were sick, with partially or fully penetrant septin defects. Genotypes labeled in red were inviable. (B) DIC images of cdc12-6 shs1 mutants (see A) at 24°C. Scale bar, 5 μm. (C) Heterozygous diploids expressing the indicated Shs1 protein fused to GFP from the SHS1 locus. Scale bar, 5 μm. (D) Scatter plot quantifying cdc12-6-SpoVM septin complex adsorption onto different membrane curvatures. Black bars represent the mean. Error bars are the SD for more than 30 measured beads at each curvature across three replicates. Adsorption of cdc12-6-SpoVM complexes was significantly greater on a membrane curvature of 2 μm−1 than on curvatures of 0.67 and 0.4 μm−1; ** (p < 0.01 and p < 0.0001, respectively). ns, adsorption was not significantly different. (E) Western blot comparing expression of indicated Cdc12 chimeras fused to 3xHA epitope in heterozygous diploids.

Article Snippet: A polyclonal rabbit α-HA epitope antibody preparation was used at 1:2000 dilution (Rockland Immunochemicals).

Techniques: Expressing, Labeling, Adsorption, Membrane, Western Blot

Journal: Development (Cambridge, England)

Article Title: GDNF availability determines enteric neuron number by controlling precursor proliferation.

doi: 10.1242/dev.00433

Figure Lengend Snippet: Fig. 3. GFRα1 and GFRα2 expression in the gut. (A,B) E14 gut expresses GFRα1 (red arrows, FITC) but not GFRα2 (FITC). Yellow arrows (B) show PGP9.5- expressing neural crest cells in the gut (Cy3 secondary), but there is no detectable GFRα2 staining. Newborn (C,D) and adult (E,F) small bowels express GFRα1 (FITC) and GFRα2 (FITC), but GFRα2 is much more easily detected in adult (F) than in newborn mouse gut. Scale bar: 100 µm.

Article Snippet: GFRα1 and GFRα2 (BAF560 and BAF429 respectively; R&D Systems, 1:40) immunohistochemistry was performed on 12 μm fresh frozen sections.

Techniques: Expressing, Staining