apc cy7 anti mouse cd45 1  (Cytek Biosciences)

Journal: Stem cells (Dayton, Ohio)

Article Title: Combination therapy with Treg and MSC enhances potency and attenuation of inflammation after traumatic brain injury compared to monotherapy

doi: 10.1002/stem.3320

Figure Lengend Snippet: (A): Representative samples from both sham and CCI groups are shown. The single cell population was identified based on SSC and FSC. Live cells were identified as negative for the 7-AAD. Microglia were identified using a two-step method. First, CD11b/c+ cells (PE-Cy7) were selected to identify all myeloid cells. The CD11b/c+ cells were then gated on P2Y12 (FITC) and CD45 (APC-Cy7). Microglia were identified as triple positive cells. SSC, side scatter; FSC, forward scatter. (B): The multiple color flow cytometry panel used to identify and immunophenotype microglia and peripheral myeloid cells.

Article Snippet: SSC, side scatter; FSC, forward scatter. (B): The multiple color flow cytometry panel used to identify and immunophenotype microglia and peripheral myeloid cells. table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Fluorochrome Antibody Clone Vendor APC-Cy7 CD45 OX-1 BD PE-Cy7 CD11b/c OX-42 BD FITC P2Y12 n/a Alomone Labs PE CD32 D34–485 BD Alexa Fluor 647 RT1B OX-6 BD PerCP Cy5.5 7-AAD n/a BD Open in a separate window Multicolor Flow Cytometry Microglia/Myeloid Cell Panel Microglia gating strategy Conventional flow cytometry analyses were performed with FlowJo vr10.6.1.

Techniques: Flow Cytometry

Journal: Stem cells (Dayton, Ohio)

Article Title: Combination therapy with Treg and MSC enhances potency and attenuation of inflammation after traumatic brain injury compared to monotherapy

doi: 10.1002/stem.3320

Figure Lengend Snippet: Multicolor Flow Cytometry Microglia/Myeloid Cell Panel

Article Snippet: SSC, side scatter; FSC, forward scatter. (B): The multiple color flow cytometry panel used to identify and immunophenotype microglia and peripheral myeloid cells. table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Fluorochrome Antibody Clone Vendor APC-Cy7 CD45 OX-1 BD PE-Cy7 CD11b/c OX-42 BD FITC P2Y12 n/a Alomone Labs PE CD32 D34–485 BD Alexa Fluor 647 RT1B OX-6 BD PerCP Cy5.5 7-AAD n/a BD Open in a separate window Multicolor Flow Cytometry Microglia/Myeloid Cell Panel Microglia gating strategy Conventional flow cytometry analyses were performed with FlowJo vr10.6.1.

Techniques: Flow Cytometry

Journal: Cell Stem Cell

Article Title: Controlling genetic heterogeneity in gene-edited hematopoietic stem cells by single-cell expansion

doi: 10.1016/j.stem.2023.06.002

Figure Lengend Snippet: Single-cell cloning of gene-edited functional HSCs (A) Schematic showing the extracellular domain of CD45 with allele-specific antibody clones 104 and A20 and the epitope-defining amino acid. (B) Experimental setup of the single-cell editing and expansion experiment. (C) Left: fractions of CD201 + CD150 + KSL cells in single-cell-derived cultures 14 days after cloning (n = 261 clones). Right: Histogram of CD201 + CD150 + KSL cell frequency. Zoomed-in region shows clones with >10% CD201 + CD150 + KSL cells. (D and E) CD45.1 + donor PB chimerism (D) and lineage distribution (E ) in single recipients with long-term (LT) engraftment ≥5% and multilineage reconstitution (n = 8). Numbers over graphs in (D) represent percentage of CD201 + CD150 + KSL cells in the transplanted clone (%). (F) Linear correlation plots of CD201 + CD150 + KSL cell frequency and 16-week donor chimerism. Red dots indicate LT repopulating and multilineage clones. Pearson correlation. (G) CD45.1 + PB chimerism and lineage distribution in secondary recipients (n = 5). See also Figure S4 and Table S3 . Error bars represent SD.

Article Snippet: anti-mouse CD45.1-APC/Cy7 (A20) , Tonbo Biosciences , Cat#25-0453; RRID: AB_2621629.

Techniques: Cloning, Functional Assay, Clone Assay, Derivative Assay

Journal: Cell Stem Cell

Article Title: Controlling genetic heterogeneity in gene-edited hematopoietic stem cells by single-cell expansion

doi: 10.1016/j.stem.2023.06.002

Figure Lengend Snippet:

Article Snippet: anti-mouse CD45.1-APC/Cy7 (A20) , Tonbo Biosciences , Cat#25-0453; RRID: AB_2621629.

Techniques: Recombinant, Sequencing, Software, CRISPR

Journal: Nature Immunology

Article Title: PI16 + reticular cells in human palatine tonsils govern T cell activity in distinct subepithelial niches

doi: 10.1038/s41590-023-01502-4

Figure Lengend Snippet: a , b , Representative flow cytometric plots depicting the gating strategy for each stromal cell type according to the indicated markers. c , Quantification of epithelial cells (EpC) as a percentage of total CD45 – CD235a – live cells in pediatric ( n = 5) and adult ( n = 6) patients with OSA. Data show average value of left and right tonsils for each patient. Mean and s.d. are indicated. P value was calculated with the two-sided Mann-Whitney test. d , Representative flow cytometric plots depicting the sorting strategy for scRNA-seq of CD45 – CD235a – stromal cells from human palatine tonsils according to the indicated markers. e – g , Expression patterns of indicated BEC, LEC and EpC gene signatures projected onto UMAPs. ScRNA-seq data represents a total of 86,966 CD45 – CD235a – stromal cells from n = 12 patients.

Article Snippet: Anti-human CD45–APC-Cy7 (clone 2D1, 560178, 1:100), anti-human CD4–BUV395 (clone SK3, 563550, 1:200), anti-human CD8–BUV805 (clone SK1, 612889, 1:400) and anti-human CD25–BUV563 (clone 2A3, 612918, 1:400) were purchased from BD Biosciences.

Techniques: MANN-WHITNEY, Expressing

Journal: Nature Immunology

Article Title: PI16 + reticular cells in human palatine tonsils govern T cell activity in distinct subepithelial niches

doi: 10.1038/s41590-023-01502-4

Figure Lengend Snippet: a , Representative UMAPs of distinct tonsillar stromal cell types according to the PhenoGraph clustering algorithm based on forward scatter (FSC)-A, side scatter (SSC)-A, CD31, ACTA2, PDPN and UEA1 flow cytometry data of CD45 − CD235a − cells ( n = 3 pediatric and n = 3 adult patients with OSA). Epithelial cells (EpCs), ACTA2 + cells, BECs, LECs, FRCs and negative cells (N) are indicated. b , UMAPs show the expression pattern of the indicated markers. c – e , Quantification of the indicated stromal cell types as a percentage of CD45 − CD235a − live cells in pediatric ( n = 5) and adult ( n = 6) patients with OSA. Data show average values of left and right tonsils for each patient. Mean and s.d. are indicated. P values were calculated with the two-sided Mann–Whitney test.

Article Snippet: Anti-human CD45–APC-Cy7 (clone 2D1, 560178, 1:100), anti-human CD4–BUV395 (clone SK3, 563550, 1:200), anti-human CD8–BUV805 (clone SK1, 612889, 1:400) and anti-human CD25–BUV563 (clone 2A3, 612918, 1:400) were purchased from BD Biosciences.

Techniques: Flow Cytometry, Expressing, MANN-WHITNEY

Journal: Nature Immunology

Article Title: PI16 + reticular cells in human palatine tonsils govern T cell activity in distinct subepithelial niches

doi: 10.1038/s41590-023-01502-4

Figure Lengend Snippet: a , Schematic representation of the scRNA-seq workflow. b , UMAP showing 14 CD45 − CD235a − stromal cell clusters after removal of contaminating cells including BECs, LECs, epithelial cells, ACTA2 + cells and FRCs. c , Dot plot depicts marker genes used for characterization and assignment of the indicated stromal cell types. Frames highlight marker genes of FRC clusters and the two ACTA2 + clusters (8 and 9). SMC, smooth muscle cell; SkMC, skeletal muscle cell. d , e , Expression patterns of VSMC and FRC gene signatures projected onto UMAPs. f , UpSet plot showing differentially expressed genes shared between VSMCs, PRCs in cluster 8 and FRCs. g , i , Top significantly enriched terms according to gene ontology (GO) enrichment analysis based on differentially expressed genes shared between the indicated cell types. Bar plots show counts of genes assigned to respective cellular processes. h , j , Expression patterns of genes assigned to the indicated cellular processes projected onto UMAPs. scRNA-seq data represent a total of 86,966 CD45 − CD235a − stromal cells from n = 12 patients. ECM, extracellular matrix.

Article Snippet: Anti-human CD45–APC-Cy7 (clone 2D1, 560178, 1:100), anti-human CD4–BUV395 (clone SK3, 563550, 1:200), anti-human CD8–BUV805 (clone SK1, 612889, 1:400) and anti-human CD25–BUV563 (clone 2A3, 612918, 1:400) were purchased from BD Biosciences.

Techniques: Marker, Expressing

Journal: Nature Immunology

Article Title: PI16 + reticular cells in human palatine tonsils govern T cell activity in distinct subepithelial niches

doi: 10.1038/s41590-023-01502-4

Figure Lengend Snippet: a – c , UMAPs display scRNA-seq data of tonsillar CD45 − CD235a − stromal cells separated and colored according to different conditions. FRCs showing strong transcriptional changes are highlighted by arrows. d , Bar plot shows the relative abundance of FRCs among CD45 − CD235a − stromal cells according to different conditions as revealed by scRNA-seq. w/o, without. e , UMAP visualization of re-embedded FRC subsets. FDC, follicular dendritic cells. f , Feature UMAPs show expression pattern of cluster marker genes used for characterization of the indicated FRC subsets. g , Chord diagram shows the proportion of cells derived from different conditions for each FRC subset. h , Diffusion map dimensionality reduction of FRC subsets. i , Top significantly enriched GO terms in PI16 + RCs according to enrichment analysis based on subset marker genes. Expression pattern of genes assigned to the indicated cellular processes is projected onto diffusion maps. BMP, bone morphogenic protein. scRNA-seq data represent a total of 86,966 CD45 − CD235a − stromal cells and 28,571 FRCs containing 20,158 cells (4,867 FRCs) from n = 3 pediatric patients with OSA, 21,596 cells (6,184 FRCs) from n = 5 adult patients with OSA and 45,212 cells (17,520 FRCs) from n = 4 adult patients with tonsillitis.

Article Snippet: Anti-human CD45–APC-Cy7 (clone 2D1, 560178, 1:100), anti-human CD4–BUV395 (clone SK3, 563550, 1:200), anti-human CD8–BUV805 (clone SK1, 612889, 1:400) and anti-human CD25–BUV563 (clone 2A3, 612918, 1:400) were purchased from BD Biosciences.

Techniques: Expressing, Marker, Derivative Assay, Diffusion-based Assay

Journal: Nature Immunology

Article Title: PI16 + reticular cells in human palatine tonsils govern T cell activity in distinct subepithelial niches

doi: 10.1038/s41590-023-01502-4

Figure Lengend Snippet: a , Bar plots show the relative abundance of indicated stromal cell types among CD45 – CD235a – cells of individual patients according to different conditions and based on scRNA-seq data. Mean and SEM are indicated. P values as per Kruskal-Wallis test. ScRNA-seq data represents a total of 86,966 CD45 – CD235a – stromal cells containing 20,158 cells from n = 3 pediatric patients with OSA, 21,596 cells from n = 5 adult patients with OSA and 45,212 cells from n = 4 adult patients with tonsillitis. b , UMAP showing 126,320 CD45 – CD235a – stromal cells from 12 patients acquired at KSSG (Kantonsspital St.Gallen, St.Gallen, Switzerland) and 3 patients independently acquired at UPENN (University of Pennsylvania, Philadelphia, USA) (see Extended Data Table ) integrated over their origin. BECs, LECs, EpCs, ACTA2 + cells and PI16 + RCs as a fraction of FRCs are indicated. c , UMAP visualization of tonsillar stromal cells split by sample origin. d , Bar plot showing the relative abundance of PI16 + RCs among CD45 – CD235a – cells per condition and based on scRNAseq data. Data represents n = 5 (Pediatric,OSA), n = 6 (Adult,OSA) and n = 4 (Adult,Tonsillitis) individual patients. Mean and SEM are indicated. P values as per two-sided Wilcoxon test. e , Heatmap showing the average expression of marker genes used for characterization of FRC subsets. f , Featureplots visualizing the expression pattern of indicated cytokines and chemokines across FRCs. g , UMAP depicting re-embedded FRCs colored according to the indicated patient groups. h , Significantly enriched terms according to GO enrichment analysis based on differentially expressed genes for the indicated FRC subsets. e-h , ScRNA-seq data represents 28,571 FRCs containing 4,867 cells from n = 3 pediatric patients with OSA, 6,184 cells from n = 5 adult patients with OSA and 17,520 cells from n = 4 adult patients with tonsillitis.

Article Snippet: Anti-human CD45–APC-Cy7 (clone 2D1, 560178, 1:100), anti-human CD4–BUV395 (clone SK3, 563550, 1:200), anti-human CD8–BUV805 (clone SK1, 612889, 1:400) and anti-human CD25–BUV563 (clone 2A3, 612918, 1:400) were purchased from BD Biosciences.

Techniques: Expressing, Marker