Journal: Cell reports

Article Title: Activation of GPR44 decreases severity of myeloid leukemia via specific targeting of leukemia initiating stem cells

doi: 10.1016/j.celrep.2023.112794

Figure Lengend Snippet:

Article Snippet: Mouse anti-mouse CD45.1 (FITC) , Miltenyi Biotec , Cat# 130–124-211; RRID: AB_2857674.

Techniques: Virus, Recombinant, Enzyme-linked Immunosorbent Assay, Binding Assay, Staining, Modification, Saline, Concentration Assay, Over Expression, Protein Extraction, Membrane, SYBR Green Assay, Bicinchoninic Acid Protein Assay, Protease Inhibitor, CCK-8 Assay, Reverse Transcription, Selection, Plasmid Preparation, Knock-Out, Software, Real-time Polymerase Chain Reaction, Flow Cytometry

Journal: eLife

Article Title: Distinct impact of IgG subclass on autoantibody pathogenicity in different IgG4-mediated diseases

doi: 10.7554/eLife.76223

Figure Lengend Snippet: ( A ) Plots showing the unit values of indicated Dsg1-specific antibodies in the serum of pemphigus foliaceus (PF) patients (n=53). ( B ) Plots showing the levels of IgG1 and IgG4 antibodies in two PF patients that bind to Dsg1, Dsg2, or Dsg1/Dsg2 chimeric molecules containing the indicated Dsg1 EC domains. ( C ) Representative photos and scores showing the hematoxylin-eosin (HE) staining results of ears of hFCGR-Tg and FcγRα-null mice 3 days after being treated with 0.4 mg of Ctrl hIgG (n=3), or PF24-9(IgG1) or PF24-9(IgG4) (n≥4) through tail vein injection. Scale bars: 200 μm. ( D–F ) Representative flow cytometry profiles ( D ) and plots showing the percentage ( E ) and cell number ( F ) of infiltrating neutrophils (CD11b + Gr1 hi ) among leukocytes (CD45 + ) in the ears of mice in ( C ). Each symbol is derived from an individual PF patient ( A ) or an individual mouse ( C, E, F ). Mean ± SEM values are plotted ( A, C, E, F ). One-way ANOVA ( A ) and two-way ANOVA ( C, E, F ) with Tukey’s multiple comparisons test. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. A representative of two independent experiments is shown. Figure 4—source data 1. Related to . Figure 4—source data 2. Related to . Figure 4—source data 3. Related to . Figure 4—source data 4. Related to . Figure 4—source data 5. Related to . Figure 4—source data 6. Related to .

Article Snippet: Antibody , Anti-mouse CD45.2 Alexa Fluor 700 (Mouse monoclonal) , BD , Cat# 560693 RRID: AB_1727491 , FACS (1:200).

Techniques: Staining, Injection, Flow Cytometry, Derivative Assay

Journal: eLife

Article Title: Distinct impact of IgG subclass on autoantibody pathogenicity in different IgG4-mediated diseases

doi: 10.7554/eLife.76223

Figure Lengend Snippet: Representative flow cytometry profile ( A ) and plots showing the percentage ( B ) and cell number ( C ) of infiltrating CD11b + myeloid cells among leukocytes (CD45 + ) in the ear tissues of mice in . Each photo or symbol is derived from an individual mouse. Mean ± SEM values are plotted. Two-way ANOVA with Tukey’s multiple comparisons test ( B, C ). *p<0.05, **p<0.01, ****p<0.0001. A representative of two independent experiments is shown.

Article Snippet: Antibody , Anti-mouse CD45.2 Alexa Fluor 700 (Mouse monoclonal) , BD , Cat# 560693 RRID: AB_1727491 , FACS (1:200).

Techniques: Flow Cytometry, Derivative Assay

Journal: eLife

Article Title: Distinct impact of IgG subclass on autoantibody pathogenicity in different IgG4-mediated diseases

doi: 10.7554/eLife.76223

Figure Lengend Snippet: ( A ) Representative photos and scores showing the hematoxylin-eosin (HE) staining results of ears of hFCGR-Tg mice 3 days after being treated with 0.5 mg of Ctrl hIgG (n=3), or PF24-9(N297A) or PF24-9(GASDALIE) autoantibodies (n=5). Scale bars: 200 μm. ( B, C ) Ear thickness ( B ) and weight ( C ) of hFCGR-Tg mice in ( A ) when sacrificed at day 3. ( D, E ) Plots showing the percentage ( D ) and cell number ( E ) of infiltrating neutrophils (CD11b + Gr1 hi ) among leukocytes (CD45 + ) in the ears of mice in ( A ). ( F ) Photos and gross phenotype scores of nude mice 2 days after being treated with 0.4 mg of Ctrl hIgG or PF1-8-15(N297A) or PF1-8-15(GASDALIE) antibodies (n=5). Each symbol is derived from an individual mouse. Mean ± SEM values are plotted. One-way ANOVA with Tukey’s multiple comparisons test ( A–F ). *p<0.05, **p<0.01, ****p<0.0001. Figure 5—source data 1. Related to . Figure 5—source data 2. Related to . Figure 5—source data 3. Related to . Figure 5—source data 4. Related to . Figure 5—source data 5. Related to . Figure 5—source data 6. Related to .

Article Snippet: Antibody , Anti-mouse CD45.2 Alexa Fluor 700 (Mouse monoclonal) , BD , Cat# 560693 RRID: AB_1727491 , FACS (1:200).

Techniques: Staining, Derivative Assay

Journal: eLife

Article Title: Distinct impact of IgG subclass on autoantibody pathogenicity in different IgG4-mediated diseases

doi: 10.7554/eLife.76223

Figure Lengend Snippet: ( A, B ) Representative hematoxylin-eosin (HE) results and scores of the ear tissues of hFCGR-Tg mice 3 days ( A ) or 6 ( B ) days after being treated with 0.5 mg of Ctrl hIgG (n=4), PF1-8-15(GASDALIE), or PF1-8-15(N297A) (n=5). Scale bars: 200 μm. ( C, D ) Ear thickness ( C ) and weight ( D ) of mice in ( B ). ( E–G ) Representative flow cytometry profiles ( E ) and plots showing the percentage ( F ) and cell number ( G ) of infiltrating neutrophils (CD11b + Gr1 hi ) among leukocytes (CD45 + ) in the ears of mice in ( B ). ( H ) Plots showing the levels of the remaining human IgG in the serum of mice in ( A ). ( I ) Photos and gross phenotype scores of nude mice 4 days after being treated with 0.5 mg of Ctrl hIgG, PF1-8-15(N297A), or PF1-8-15(GASDALIE) (n=5). Each symbol is derived from an individual mouse. Mean ± SEM values are plotted. One-way ANOVA with Tukey’s multiple comparisons test ( A–D, F–I ). *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. A representative of three independent experiments is shown.

Article Snippet: Antibody , Anti-mouse CD45.2 Alexa Fluor 700 (Mouse monoclonal) , BD , Cat# 560693 RRID: AB_1727491 , FACS (1:200).

Techniques: Flow Cytometry, Derivative Assay

Journal: eLife

Article Title: Distinct impact of IgG subclass on autoantibody pathogenicity in different IgG4-mediated diseases

doi: 10.7554/eLife.76223

Figure Lengend Snippet: ( A ) Pictures of the back skin of mice treated with 0.5 mg of Ctrl hIgG or anti-Dsg1 antibody variants, photos of 3 representative mice/group (n=4 or 5) at indicated time points (day 3, day 6, day 12, and day 23) were shown. ( B ) Hematoxylin-eosin (HE) results showing the recovery of skin lesions and inflammation on day 23. Scale bars: 200 μm. ( C–D ) Plots showing the percentage ( C ) and cell number ( D ) of infiltrating neutrophils (CD11b + Gr1 hi ) among leukocytes (CD45 + ) in the ears of mice 23 days after antibodies treatment. ( E ) Plots showing the levels of the remaining human IgG in the serum of mice at the indicated time points. ( F ) Direct immunofluorescence results showing the clearance of human IgG binding to Dsg1 in skin tissue. Hu Tonsi is used as the positive control. Scale bars: 100 μm. Each photo or symbol is derived from an individual mouse. Mean ± SEM values are plotted. One-way ANOVA with Tukey’s multiple comparisons ( C, D ).

Article Snippet: Antibody , Anti-mouse CD45.2 Alexa Fluor 700 (Mouse monoclonal) , BD , Cat# 560693 RRID: AB_1727491 , FACS (1:200).

Techniques: Immunofluorescence, Binding Assay, Positive Control, Derivative Assay

Journal: eLife

Article Title: Distinct impact of IgG subclass on autoantibody pathogenicity in different IgG4-mediated diseases

doi: 10.7554/eLife.76223

Figure Lengend Snippet: ( A ) Representative hematoxylin-eosin (HE) results and scores of the ear tissues of hFCGR-Tg and FcγRα-null mice 3 days after being treated with 0.5 mg of Ctrl hIgG (n=2) or PF1-8-15(GASDALIE) (n=4). Scale bars: 200 μm. ( B ) Plot showing ear weight of mice in ( A ). ( C–E ) Representative flow cytometry profiles ( C ) and plots showing the percentage ( D ) and cell number ( E ) of infiltrating neutrophils (CD11b + Gr1 hi ) among leukocytes (CD45 + ) in the ears of mice in ( A ). Each symbol is derived from an individual mouse. Mean ± SEM values are plotted. Two-way ANOVA with Tukey’s multiple comparisons test. *p<0.05, **p<0.01, ****p<0.0001. A representative of two independent experiments is shown.

Article Snippet: Antibody , Anti-mouse CD45.2 Alexa Fluor 700 (Mouse monoclonal) , BD , Cat# 560693 RRID: AB_1727491 , FACS (1:200).

Techniques: Flow Cytometry, Derivative Assay

Journal: eLife

Article Title: Distinct impact of IgG subclass on autoantibody pathogenicity in different IgG4-mediated diseases

doi: 10.7554/eLife.76223

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-mouse CD45.2 Alexa Fluor 700 (Mouse monoclonal) , BD , Cat# 560693 RRID: AB_1727491 , FACS (1:200).

Techniques: Expressing, Recombinant, Enzyme-linked Immunosorbent Assay, Software, Staining

Journal: eLife

Article Title: Response to immune checkpoint blockade improved in pre-clinical model of breast cancer after bariatric surgery

doi: 10.7554/eLife.79143

Figure Lengend Snippet: ( A ) Database for annotation, visualization, and integrated discovery (DAVID) analysis of regulated pathways and false discovery rate (FDR) for high fat diet (HFD)-vertical sleeve gastrectomy (VSG) (red) and weight-matched sham (WM-Sham) (blue) relative to tumors from HFD-Sham mice is shown. N=6–8. ( B ) Heat map of row mean centered gene expression in tumor by RNA sequencing (RNA-seq) of genes contributing to significantly regulated inflammatory response pathway (GO:0006954) and response to hypoxia pathway (GO:0001666) determined by DAVID analysis. N=6–8. ( C ) Flow cytometric analysis of CD45 negative (CD45−) PD-L1+ non-immune cells in tumor is plotted as frequency of total live cells. ( D ) Mean fluorescent intensity (MFI) of PD-L1 on CD45− PD-L1+ cells in tumor is shown. N=4–5. ( E ) Circulating IL-6 concentration in plasma was measured at endpoint after 4 hr of fasting by Luminex. N=8–14. ( F ) Flow cytometric analysis of PD-L1 MFI in E0771 breast cancer cells after treatment with recombinant mouse IL-6 (200 pg/mL) for 4 hr. Mean ± SEM is shown. One-way ANOVA with Fisher’s LSD test. *p<0.05, **p<0.01, and ***p<0.001. ( G ) Gene set enrichment analysis (GSEA) of the hallmark pathway for IL6/JAK/STAT3 gene set from the Molecular Signatures Database of the Broad Institute is reported in HFD-VSG tumors compared to WM-Sham controls. The normalized enrichment score (NES) and FDR are shown.

Article Snippet: Antibody , Anti-mouse CD45 violetFluor 450 (Rat monoclonal) , Tonbo Biosciences , Cat# 75–0451 U025 , (1:40).

Techniques: Gene Expression, RNA Sequencing, Concentration Assay, Clinical Proteomics, Luminex, Recombinant

Journal: eLife

Article Title: Response to immune checkpoint blockade improved in pre-clinical model of breast cancer after bariatric surgery

doi: 10.7554/eLife.79143

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-mouse CD45 violetFluor 450 (Rat monoclonal) , Tonbo Biosciences , Cat# 75–0451 U025 , (1:40).

Techniques: Transfection, Luciferase, Recombinant, Sequencing, Control

Journal: eLife

Article Title: Polycomb repressive complex 1.1 coordinates homeostatic and emergency myelopoiesis

doi: 10.7554/eLife.83004

Figure Lengend Snippet:

Article Snippet: Antibody , violetFluor 450 anti-mouse CD45.2 (104) (mouse monoclonal) , Tonbo Biosciences , Cat# 75-0454; RRID :AB_2621950 , FACS (4 × 10 –2 μL/1 × 10 6 cell).

Techniques: Produced, Sequencing, Recombinant, Protease Inhibitor, Purification, Staining, Avidin-Biotin Assay, Blocking Assay, Plasmid Preparation, Software, Immunostaining

Journal: Bone Marrow Transplantation

Article Title: Brief ex vivo Fas-ligand incubation attenuates GvHD without compromising stem cell graft performance

doi: 10.1038/s41409-020-0941-2

Figure Lengend Snippet: a – h MPBC graft characterization following FasL treatment. Percentage of annexin V positive a CD3 + and b CD34 + cells. c Percentage of CD3 + and d CD34 + cells per total CD45 + population. HSPCs subpopulations; e Immature (CD34 + CD38 low ), f Multipotent progenitors (CD45RA − CD90 − ) and g self-renewing hematopoietic stem cells (CD45RA − CD90 + ). h Colony-forming units (CFU) profile of: erythroid progenitor cells (CFU-E and BFU-E), granulocyte-macrophage progenitor cells (CFU-GM) and multipotential granulocyte, erythroid, macrophage, megakaryocyte progenitor cells (CFU-GEMM). Engraftment, differentiation and CFU potential as detected in the BM of γ-irradiated (2.75 Gy) NSG mice, 4 weeks post transplantation of 1 × 10 5 human CD34 + cells: i human leukocytes (hCD45 + ) j immature hCD34 + CD38 low progenitors and k human leukocytes subpopulations: B (hCD19 + ), Myelo-monocytic (hCD33 + and CD14 + CD16 − ), NK (hCD56 + CD16 − ), HSPCs (CD34 + ) cells and l number of human colony-forming cells in the mice BM. Data presented as ( a – h ) mean+SD or ( i – l ) individual mice and median. ( a – d ) n = 25 or ( e – h ) n = 3 individual MPBCs donations and ( i – l ) show one representative study out of two, n = 7 (MPBC-CD34) and n = 8 (MPBC + FasL-CD34) female mice per group transplanted with cells from one individual MPBC donation. Statistical analysis preformed using ( a – d ) ANOVA parametric test, ( e – h ) paired T-test or ( i – l ) Mann–Whitney test. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The following antibodies were used: anti- mouse CD45 (130-102-412), human CD45 (130-098-143, 130-098-151, 130-098-141, 130-098-148, 130-104-566, 130-110-637), CD34 (130-081-002, 130-090-954), CD90 (130-099-289), CD3 (130-109-460, 130-109-466), CD4 (130-110-680), CD8 (130-109-454), CCR7 (130-108-309), CD45RA (130-108-784, 130-110-637), LFA1 (130-105-437), CD95 (130-104-232), CXCR3 (130-101-378), CCR6 (130-100-377), CD25 (130-109-021), CD33 (130-111-021), CD19 (130-113-642, 130-110-249), CD27 (130-099-499), CD38 (130-108-862), HLA-DR (130-104-825), IL17 (130-100-077), IFN-γ (130-097-944) and FoxP3 (130-098-119) all purchased from Miltenyi Biotech (Bergisch Gladbach, Germany); matched isotype controls were used as negative control.

Techniques: Irradiation, Transplantation Assay, MANN-WHITNEY

Journal: Molecular Neurodegeneration

Article Title: Prevention of C5aR1 signaling delays microglial inflammatory polarization, favors clearance pathways and suppresses cognitive loss

doi: 10.1186/s13024-017-0210-z

Figure Lengend Snippet: Fibrillar plaque and CD45 microglial expression in hippocampus. Representative pictures of thioflavine-S staining (fibrillar plaques) (top panels) and CD45 staining (microglia) (lower panels) at 7 ( a ) and 10 months ( c ). Scale bar: 50 um. ( b ) Image analysis of 7 month thioflavine-S (upper) and CD45 (lower) staining (Arctic n = 9, Arctic/C5aR1KO n = 9) ( d ) Image analysis of 10 mo thioflavine (Arctic n = 12, Arctic/C5aR1KO n = 9) (upper) and CD45 (Arctic n = 11, Arctic/C5aR1KO n = 7) lower). Bars represent group means +/− SEM of n animals per genotype. Data represent the average of 2 to 4 sections per animal. P value was calculated using one- way ANOVA

Article Snippet: Tissues were incubated overnight at 4 °C with anti mouse CD45 antibodies (rat monoclonal anti CD45, 1 μg/mL, Serotec, or goat anti CD45 0.2 μg/mL, R&D systems, Minneapolis, MN), anti Aß antibody (rabbit polyclonal 1536 [ ] gift from N.R Cooper) or appropriate control IgG in blocking solution.

Techniques: Expressing, Staining

Journal: eLife

Article Title: Integrative analysis of scRNA-seq and scATAC-seq revealed transit-amplifying thymic epithelial cells expressing autoimmune regulator

doi: 10.7554/eLife.73998

Figure Lengend Snippet: ( A ) Uniform manifold approximation and production (UMAP) plot of scATAC-seq data from TECs (EpCAM + CD45 – TER119 – ) from 4-week mice. Cell clusters are separated by colors and numbers in the plot. The two datasets were integrated using the Seurat package. The graph on the right shows percentages of each cluster in the total number of cells detected (15,255 cells). ( B ) Chromatin accessibility of typical marker genes of TECs. Accessibility in each gene region is represented in red. ( C ) Violin plot depicting chromatin accessibility in Aire and Cd80 gene regions in each cluster. ( D ) Pseudo-bulk accessibility tracks of the Aire gene region in each cluster (upper panels) and frequency of sequenced fragments within the Aire gene region of individual cells in cluster 0, 2, and 4 (lower panels).

Article Snippet: Antibody , APC/Cyanine7 anti-mouse CD45(Rat monoclonal) , BioLegend , Cat#103116, RRID: AB_312981 , FACS (1:200).

Techniques: Marker

Journal: eLife

Article Title: Integrative analysis of scRNA-seq and scATAC-seq revealed transit-amplifying thymic epithelial cells expressing autoimmune regulator

doi: 10.7554/eLife.73998

Figure Lengend Snippet: ( A ) Uniform manifold approximation and production (UMAP) plot of scRNA-seq data from TECs (EpCAM + CD45 – TER119 – ) from 4-week mice. Cell clusters (R0 to R17) are indicated by colors and numbers in the plot. The graph on the right shows the percentages of each cluster in the total number of cells detected (11,792 cells). ( B ) Violin plots depicting expression levels of typical TEC marker genes in each cluster.

Article Snippet: Antibody , APC/Cyanine7 anti-mouse CD45(Rat monoclonal) , BioLegend , Cat#103116, RRID: AB_312981 , FACS (1:200).

Techniques: Expressing, Marker

Journal: eLife

Article Title: Integrative analysis of scRNA-seq and scATAC-seq revealed transit-amplifying thymic epithelial cells expressing autoimmune regulator

doi: 10.7554/eLife.73998

Figure Lengend Snippet:

Article Snippet: Antibody , APC/Cyanine7 anti-mouse CD45(Rat monoclonal) , BioLegend , Cat#103116, RRID: AB_312981 , FACS (1:200).

Techniques: Transgenic Assay, Purification, Plasmid Preparation, Staining, Software, Multiplex Assay