Journal: PLoS ONE

Article Title: Nogo-Receptor 1 Deficiency Has No Influence on Immune Cell Repertoire or Function during Experimental Autoimmune Encephalomyelitis

doi: 10.1371/journal.pone.0082101

Figure Lengend Snippet: ( A ) Comparative flow cytometric analysis of single cell suspension from spleen, BM, thymus, blood, lymph nodes and central nervous system (CNS) associated mononuclear cells. Proportion and total number of CD3 + CD8 + and CD3 + CD4 + T cells, B220 + B cells, Gr-1 + granulocytes and F4/80 (Gr-1 lo F4/80 + ) monocyte/macrophage are shown. Data represent mean ± SEM (n = 8-11). *p<0.05 Mann-Whitney test. ( B ) Quantification of BM-derived colony number in naïve ngr1-/- and WT mice . After 8 days on methylcellulose culture, BM-derived progenitors from ngr1-/- animals were capable of producing granulocytes (G); macrophages (M) and mixed (GM) colonies at comparable numbers to WT mice. Bars represent mean colony number/plate ± SEM (n = 3).

Article Snippet: Single cell suspensions were prepared and 1×10 5 mononuclear cells were seeded in triplicate in methylcellulose medium (Stem Cell Technologies, Vancouver, Canada) suitable for the growth of granulocyte-macrophage progenitors.

Techniques: Suspension, MANN-WHITNEY, Derivative Assay

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: Cell reports

Article Title: Autism-associated chromatin remodeler CHD8 regulates erythroblast cytokinesis and fine-tunes the balance of Rho GTPase signaling

doi: 10.1016/j.celrep.2022.111072

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Serum-free methylcellulose-based medium with recombinant cytokines (including EPO) for mouse erythroid progenitor cells , STEMCELL Technologies , Cat # 03436.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Staining, Diagnostic Assay, Sample Purification, Multiplex Assay, RNA Sequencing, Sequencing, Real-time Polymerase Chain Reaction, Software