Journal: Blood

Article Title: A topological view of human CD34 + cell state trajectories from integrated single-cell output and proteomic data

doi: 10.1182/blood-2018-10-878025

Figure Lengend Snippet: Functionally defined progenitors with NM, L, and/or E potential display partially overlapping molecular profiles. (A-B) Distribution of each progenitor type in t-SNE space. Index sort information was used to map each progenitor to its nearest 10 neighbors in the mass cytometry data based on the scaled intensity of expression of CD45RA, CD71, CD45, CD123, CD34, CD33, CD49f, CD10, CD135, CD38, CD90, HLA-DR, and CD133. The nearest neighbors for all members of a given progenitor type were pooled and used to generate a probability density, indicated by the intensity of the color shown. The lowest level contains 95% of the total probability density, with each higher 10% density levels indicated thereafter. The black contour shows the 75th quantile of the overall density. (A) Mappings for all progenitor types assessed visually in methylcellulose assays as shown in Figure 1B. (B) Mappings for a representative selection of lineage competencies assessed in the STC assays as shown in Figure 1C. (C) A hierarchical clustering of the progenitor types analyzed based on a pairwise assessment of differences in the density distributions between all mappings of functionally and phenotypically defined cell types. Closely related groups are highlighted and given a descriptive name. (D) Multidimensional scaling indicating the relative distances (based on the distribution differences) between all phenotypically and functionally defined progenitor subsets.

Article Snippet: In vitro assays Colony formation in methylcellulose was assessed by single, randomly selected, index-sorted cells deposited directly and individually into the 60 inner wells of a flat bottom Nunc 96-well polystyrene plate (Thermo Fisher Scientific, Waltham, MA) preloaded with 50 µL of methylcellulose medium; the medium was supplemented with 50 ng/mL stem cell factor (SCF), 20 ng/mL granulocyte-macrophage colony-stimulating factor, interleukin-3 (IL-3), interleukin-6, granulocyte-colony stimulating factor (G-CSF), and 3 U/mL erythropoietin (EPO) (STEMCELL Technologies).

Techniques: Mass Cytometry, Expressing, Selection

Journal: Histology and histopathology

Article Title: Lymphoid hyperplasia and lymphoma in KSHV K1 transgenic mice

doi: 10.14670/HH-30.559

Figure Lengend Snippet: (A) Hematoxylin/eosin (H&E) staining and immunohistochemistry staining of K1and kappa/lambda light chains in spleens from one control (K1-negative littermate) and two K1 transgenic mice; notable are clusters of kappa-positive and scattered single lambda-positive cells; (B) IL-6 (left panel) and CXCR5 (right panel) mRNA levels were analyzed by qRT-PCR in splenic cells isolated from control WT (n=2) and K1-transgenic (n=10) mice; 2 control samples were pooled together; IL-6 average level 4.07 relative to controls (range 1.15 – 7.16); CXCR5 average level 1.24 relative to controls (range 0.62–1.90); (C) Splenocytes from 6 month old K1 transgenic mice and control K1-negative littermates were harvested and incubated with 50 ng/mL of anti-Fas antibody Jo2 for the indicated times. Subsequently, cells were analyzed for apoptosis; (D) K1 transgenic mice and their K1-negative littermates were challenged with a lethal dose of anti-Fas antibody Jo2 and monitored for survival up to 6 hours post challenge.

Article Snippet: Cells were resuspended in RPMI medium supplemented with 10% fetal bovine serum (both from HyClone, Logan, UT), and aliquots of 1 × 10 6 cells per mL were incubated 50 ng/mL of agonistic anti-mouse Fas antibody Jo2 (BD Pharmingen, San Jose, CA) crosslinked with 2 μg/mL of protein G (Invitrogen– Life Technologies, Carlsbad, CA) to induce apoptosis.

Techniques: Staining, Immunohistochemistry, Transgenic Assay, Quantitative RT-PCR, Isolation, Incubation