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anti human leptin receptor antibody (Miltenyi Biotec)

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Miltenyi Biotec anti human leptin receptor antibody
Anti Human Leptin Receptor Antibody, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 92/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/lepr+pe/CD295+(LEPR)+Antibody%2C+anti-human%2C+PE%2C+REAfinity/pm36759597-222-2-9
Average 92 stars, based on 7 article reviews

anti human leptin receptor antibody - by Bioz Stars, 2026-09

92/100 stars

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FACS:

Article Title: Obesity-Associated Hypermetabolism and Accelerated Senescence of Bone Marrow Stromal Stem Cells Suggest a Potential Mechanism for Bone Fragility.
Article Snippet: The flow cytometry was performed by BD LSR II (BD Biosciences) and analyzed by Kaluza 1.1 analysis software. .. For the cell sorting experiments, hBM-MSCswere incubated with Fc-blocking solution followed by incubation with pre-conjugated antibodies either LEPR-PE (Miltenyi) or IR- PE antibody (BD Bioscience) and were run through a BD FACS Aria III (BD Bioscience). ..

Incubation:

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A ) UMAP computed from 19,257 mesenchymal cells from 12 individuals showing different mesenchymal subsets, with RNAlo MSCs being excluded. B ) Dot plot showing normalized expression of key mesenchymal marker genes in MSC subsets. Rows and columns were manually ordered. C ) Dot plot showing the normalized expression of literature-derived marker genes for human MSCs including NT5E (CD73), THY1 (CD90), and ENG (CD105). NGFR (CD271) and MCAM (CD146) have also been described as canonical MSC markers. D ) CytoTRACE analysis projected onto the MSC UMAP showing the predicted differentiation score, where higher values imply the cell is more primitive. E) Boxplots showing the relative colony forming potential (number of colonies produced by each cell type divided by total colonies from each sample, each data point is one sample) of sorted Fibro-MSC (CD45-CD38-CD235ab-VECAD-PDPN+), THY1+ MSC (CD45-CD38-CD235ab-VECAD-PDPN-LEPR+ CD90+), Adipo-MSC (CD45-CD38-CD235ab-VECAD-PDPN-LEPR+ CD90-), and osteolineage cells (CD45-CD38-CD235ab-VECAD-PDPN-CD56+). P-values were computed as Fibro-MSC vs. all by Dunnett’s Multiple Comparisons test. F) Line plot showing the population doublings over the course of cell culturing for eight passages, where all cells were passaged every 7 days. P-values are computed as Fibro-MSC vs other MSCs by two-way ANOVA. G ) UMAP showing 3,874 endothelial cells from the 12 individuals. SEC – Sinusoidal Endothelial Cells, AEC – Arterial Endothelial Cells. H ) Dot plot showing normalized expression of top differentially expressed genes by adjusted p-value between endothelial subsets, as well as CDH5 , CD34 , and KDR which were manually selected as pan-endothelial markers. I ) MSCs from human bone marrow aspirate reported from De Jong et al., Nature Immunology , 2021 were reference mapped to our scRNA-Seq atlas and the cell type labels were predicted. J ) Human fetal bone marrow mesenchymal cells from Jardine et al., Nature , 2021 were reference mapped to our scRNA-Seq atlas and the cell labels were predicted.

Journal: bioRxiv

Article Title: Mapping the Cellular Biogeography of Human Bone Marrow Niches Using Single-Cell Transcriptomics and Proteomic Imaging

doi: 10.1101/2024.03.14.585083

Figure Lengend Snippet: A ) UMAP computed from 19,257 mesenchymal cells from 12 individuals showing different mesenchymal subsets, with RNAlo MSCs being excluded. B ) Dot plot showing normalized expression of key mesenchymal marker genes in MSC subsets. Rows and columns were manually ordered. C ) Dot plot showing the normalized expression of literature-derived marker genes for human MSCs including NT5E (CD73), THY1 (CD90), and ENG (CD105). NGFR (CD271) and MCAM (CD146) have also been described as canonical MSC markers. D ) CytoTRACE analysis projected onto the MSC UMAP showing the predicted differentiation score, where higher values imply the cell is more primitive. E) Boxplots showing the relative colony forming potential (number of colonies produced by each cell type divided by total colonies from each sample, each data point is one sample) of sorted Fibro-MSC (CD45-CD38-CD235ab-VECAD-PDPN+), THY1+ MSC (CD45-CD38-CD235ab-VECAD-PDPN-LEPR+ CD90+), Adipo-MSC (CD45-CD38-CD235ab-VECAD-PDPN-LEPR+ CD90-), and osteolineage cells (CD45-CD38-CD235ab-VECAD-PDPN-CD56+). P-values were computed as Fibro-MSC vs. all by Dunnett’s Multiple Comparisons test. F) Line plot showing the population doublings over the course of cell culturing for eight passages, where all cells were passaged every 7 days. P-values are computed as Fibro-MSC vs other MSCs by two-way ANOVA. G ) UMAP showing 3,874 endothelial cells from the 12 individuals. SEC – Sinusoidal Endothelial Cells, AEC – Arterial Endothelial Cells. H ) Dot plot showing normalized expression of top differentially expressed genes by adjusted p-value between endothelial subsets, as well as CDH5 , CD34 , and KDR which were manually selected as pan-endothelial markers. I ) MSCs from human bone marrow aspirate reported from De Jong et al., Nature Immunology , 2021 were reference mapped to our scRNA-Seq atlas and the cell type labels were predicted. J ) Human fetal bone marrow mesenchymal cells from Jardine et al., Nature , 2021 were reference mapped to our scRNA-Seq atlas and the cell labels were predicted.

Article Snippet: Isolated cells were stained for 15-30 minutes using anti-CD90-Bv650(Clone 5E10), anti-CD38-Bv785 (Clone HIT2), anti-CD45-FITC(Clone HI30), anti-LEPR-PE (Clone # 52263,R&D Systems), anti-Podoplanin-PE/Dazzle (Clone NC-08), anti-CD56-PE/Cy7 (Clone 5.1H11), anti-VE-Cadherin-Alexa Fluor 647 (Clone BV9), and anti-CD235ab-APC/Fire (Clone HIR2).

Techniques: Expressing, Marker, Derivative Assay, Produced, Cell Culture

Representative confocal images of VE-cadherin + (magenta) and CD31 + (green) BM vessels, and nuclei (blue) in femurs from a non-IRR mice and at b day +3 and day +7 following 500 cGy and treatment with IgG (10 µg/dose) or anti-NRP1 (10 µg/dose). Left image shows full view taken using 20× lens and right images display individual channels from magnified yellow box. Scale bar 50 µm; magnified view scale bar, 20 µm. c Quantification of VE-Cadherin vascular area from the images in a , b . Dotted line represents mean Non-IRR vascular area. Data presented as mean values +/− SEM. ( n = 2 independent experiments, d3 IgG n = 4 fields of view, d3 anti-NRP1 n = 5 fields, d7 IgG n = 7 fields, d7 anti-NRP1 n = 7 fields; IgG d3: p = 0.016, anti-NRP1 d3: p = 0.0467, IgG d7: p = 0.0349, anti-NRP1 d7: p = 0.9694. d BM cell counts at day +7 following 500 cGy and treatment with IgG or anti-NRP1 ( n = 5 mice/group, p = 0.0025). Dotted line shows cell counts of non-IRR BM. e At left, representative flow cytometric analysis of CD45 − VE-cad + BM ECs within BM lin − cells from the groups shown. At right, percentages of BM ECs at day +7 following 500 cGy. Dotted line represents %VE-cad + ECs in non-IRR controls ( n = 5 mice/group). f At left, representative histograms of % activated caspase 3/7 + cells within lin − CD45 − VE-cad + BM ECs at day +7 following 500 cGy. At right, mean % activated caspase 3/7 + BM ECs at day +7 following 500 cGy ( n = 5/group). Dotted line represents % activated caspase 3/7 + ECs in non-IRR controls. g Levels of Evans Blue Dye (EBD) in the BM extracellular space at +24 h following 500 cGy and treatment with IgG or anti-NRP1 ( n = 5–6 mice/group, non-IRR vs. IgG: p = 0.0024, IgG vs. anti-NRP1: p = 0.188, non-IRR vs. anti-NRP1 p = 0.2679). d – f Data assessed by Student’s two-tailed t -test, g One-way ANOVA with Holm-Sidak’s multiple comparison two-sided t -test, c Two-sided one-sample t -test and Wilcoxon test used to compare sample means to the mean of non-IRR samples, * p < 0.05, ** p < 0.01, **** p < 0.0001. Source data provided as a Source Data file.

Journal: Nature Communications

Article Title: Neuropilin 1 regulates bone marrow vascular regeneration and hematopoietic reconstitution

doi: 10.1038/s41467-021-27263-y

Figure Lengend Snippet: Representative confocal images of VE-cadherin + (magenta) and CD31 + (green) BM vessels, and nuclei (blue) in femurs from a non-IRR mice and at b day +3 and day +7 following 500 cGy and treatment with IgG (10 µg/dose) or anti-NRP1 (10 µg/dose). Left image shows full view taken using 20× lens and right images display individual channels from magnified yellow box. Scale bar 50 µm; magnified view scale bar, 20 µm. c Quantification of VE-Cadherin vascular area from the images in a , b . Dotted line represents mean Non-IRR vascular area. Data presented as mean values +/− SEM. ( n = 2 independent experiments, d3 IgG n = 4 fields of view, d3 anti-NRP1 n = 5 fields, d7 IgG n = 7 fields, d7 anti-NRP1 n = 7 fields; IgG d3: p = 0.016, anti-NRP1 d3: p = 0.0467, IgG d7: p = 0.0349, anti-NRP1 d7: p = 0.9694. d BM cell counts at day +7 following 500 cGy and treatment with IgG or anti-NRP1 ( n = 5 mice/group, p = 0.0025). Dotted line shows cell counts of non-IRR BM. e At left, representative flow cytometric analysis of CD45 − VE-cad + BM ECs within BM lin − cells from the groups shown. At right, percentages of BM ECs at day +7 following 500 cGy. Dotted line represents %VE-cad + ECs in non-IRR controls ( n = 5 mice/group). f At left, representative histograms of % activated caspase 3/7 + cells within lin − CD45 − VE-cad + BM ECs at day +7 following 500 cGy. At right, mean % activated caspase 3/7 + BM ECs at day +7 following 500 cGy ( n = 5/group). Dotted line represents % activated caspase 3/7 + ECs in non-IRR controls. g Levels of Evans Blue Dye (EBD) in the BM extracellular space at +24 h following 500 cGy and treatment with IgG or anti-NRP1 ( n = 5–6 mice/group, non-IRR vs. IgG: p = 0.0024, IgG vs. anti-NRP1: p = 0.188, non-IRR vs. anti-NRP1 p = 0.2679). d – f Data assessed by Student’s two-tailed t -test, g One-way ANOVA with Holm-Sidak’s multiple comparison two-sided t -test, c Two-sided one-sample t -test and Wilcoxon test used to compare sample means to the mean of non-IRR samples, * p < 0.05, ** p < 0.01, **** p < 0.0001. Source data provided as a Source Data file.

Article Snippet: For Leptin receptor+(LepR + ) stromal cells, CD45 − LepR + cells (1:200, rabbit anti-PE leptin receptor antibody, bs-0961R-PE, BiosS, Woburn, MA) were isolated.

Techniques: Two Tailed Test

a PB WBC, neutrophil (NEU), and lymphocyte (LYMPH) counts at days +3, +7, and +10 following 500 cGy and treatment with IgG or anti-NRP1 ( n = 5–10 mice/group, mean values +/− SEM; WBC: d7: p = 0.0016, d10 p = 0.002; NEU: d7: p = 0.0089, d10: p = 0.0035, LYMPH: d10: p = 0.0482). b BM cell counts at day +10 following 500 cGy and treatment with IgG or anti-NRP1 ( n = 5–10 mice/group, p = 0.0033). Dotted line represents BM cell counts in non-IRR controls. c At left, flow cytometry of BM c-kit + sca-1 − lin − progenitors and KSL cells at day +10 following 500 cGy; at right, numbers of BM c-kit + sca-1 − lin − cells and KSL at day +10 ( n = 5–10 mice/group, p < 0.0001). d At left, PB donor CD45.2 + cell engraftment at 20 weeks in CD45.1 + mice transplanted with BM cells collected at day +10 from 500 cGy—irradiated CD45.2 + mice treated with IgG or anti-NRP1, along with CD45.1 + BM competitor cells. At right, % total CD45.2 + cells, CD45.2 + Mac1 + Gr1 + myeloid cells, CD45.2 + B220 + B cells, and CD45.2 + CD3 + T cells at 20 weeks ( n = 12–13 mice/group, total: p = 0.0288, myeloid: p = 0.0266, B cells: p = 0.0051, T cells: p = 0.0001). e At left, histograms of donor CD45.2 + KSL cells in the BM of transplanted CD45.1 + mice at 20 weeks ( n = 5 mice/group, p = 0.0079). f Percent survival of C57BL/6J mice following 800 cGy TBI and treatment with anti-NRP1 or IgG for 10 days (Log-rank test, p = 0.001, n = 12 mice/group). g PB WBCs and hemoglobin (Hb) at day +7, +10 and +14 from C57BL/6J mice following 800 cGy and treatment with anti-NRP1 or IgG ( n = 8–10 mice/condition; WBC d7: p = 0.4045, WBC d10: p = 0.0415, WBC d14: p = 0.8714; Hb d7: p = 0.0154, Hb d10: p = 0.2741, Hb d14: p = 0.6216). h At left, flow cytometry of BM KSL cells 10 days following 800 cGy and treatment with anti-NRP1 or IgG; at right, %KSL cells ( n = 10 mice/condition; p = 0.0358). a Two-way ANOVA with Holm Sidak’s multiple comparison two-sided t -test. b , c Two-sided t -test, d , e Two-sided Mann–Whitney test, f Two-sided log-rank test, g Two-way ANOVA with Holm-Sidak’s multiple comparison two-sided t -tests, and h Student’s two-sided unpaired t -test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Neuropilin 1 regulates bone marrow vascular regeneration and hematopoietic reconstitution

doi: 10.1038/s41467-021-27263-y

Figure Lengend Snippet: a PB WBC, neutrophil (NEU), and lymphocyte (LYMPH) counts at days +3, +7, and +10 following 500 cGy and treatment with IgG or anti-NRP1 ( n = 5–10 mice/group, mean values +/− SEM; WBC: d7: p = 0.0016, d10 p = 0.002; NEU: d7: p = 0.0089, d10: p = 0.0035, LYMPH: d10: p = 0.0482). b BM cell counts at day +10 following 500 cGy and treatment with IgG or anti-NRP1 ( n = 5–10 mice/group, p = 0.0033). Dotted line represents BM cell counts in non-IRR controls. c At left, flow cytometry of BM c-kit + sca-1 − lin − progenitors and KSL cells at day +10 following 500 cGy; at right, numbers of BM c-kit + sca-1 − lin − cells and KSL at day +10 ( n = 5–10 mice/group, p < 0.0001). d At left, PB donor CD45.2 + cell engraftment at 20 weeks in CD45.1 + mice transplanted with BM cells collected at day +10 from 500 cGy—irradiated CD45.2 + mice treated with IgG or anti-NRP1, along with CD45.1 + BM competitor cells. At right, % total CD45.2 + cells, CD45.2 + Mac1 + Gr1 + myeloid cells, CD45.2 + B220 + B cells, and CD45.2 + CD3 + T cells at 20 weeks ( n = 12–13 mice/group, total: p = 0.0288, myeloid: p = 0.0266, B cells: p = 0.0051, T cells: p = 0.0001). e At left, histograms of donor CD45.2 + KSL cells in the BM of transplanted CD45.1 + mice at 20 weeks ( n = 5 mice/group, p = 0.0079). f Percent survival of C57BL/6J mice following 800 cGy TBI and treatment with anti-NRP1 or IgG for 10 days (Log-rank test, p = 0.001, n = 12 mice/group). g PB WBCs and hemoglobin (Hb) at day +7, +10 and +14 from C57BL/6J mice following 800 cGy and treatment with anti-NRP1 or IgG ( n = 8–10 mice/condition; WBC d7: p = 0.4045, WBC d10: p = 0.0415, WBC d14: p = 0.8714; Hb d7: p = 0.0154, Hb d10: p = 0.2741, Hb d14: p = 0.6216). h At left, flow cytometry of BM KSL cells 10 days following 800 cGy and treatment with anti-NRP1 or IgG; at right, %KSL cells ( n = 10 mice/condition; p = 0.0358). a Two-way ANOVA with Holm Sidak’s multiple comparison two-sided t -test. b , c Two-sided t -test, d , e Two-sided Mann–Whitney test, f Two-sided log-rank test, g Two-way ANOVA with Holm-Sidak’s multiple comparison two-sided t -tests, and h Student’s two-sided unpaired t -test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Source data are provided as a Source Data file.

Article Snippet: For Leptin receptor+(LepR + ) stromal cells, CD45 − LepR + cells (1:200, rabbit anti-PE leptin receptor antibody, bs-0961R-PE, BiosS, Woburn, MA) were isolated.

Techniques: Flow Cytometry, Irradiation, MANN-WHITNEY

a qRT-PCR analysis of Sema3a and Nrp1 expression in BM ECs from mice at +24 h following 5-FU chemotherapy. For each gene, expression is normalized to vehicle treatment ( n = 5/condition; Sema3a : p < 0.0001, Nrp1 : p = 0.1291). b Representative images of VE-cadherin + (magenta) , CD31 + (green) BM vessels in femur sections from untreated control mice and at day +7 following 5-FU chemotherapy and treatment with anti-NRP1 or IgG. Nuclei are stained with DAPI (blue). Scale bar, 50 µm; magnified view scale bar, 20 µm. c BM cell counts at day +7 following 5-FU chemotherapy and treatment with anti-NRP1 or IgG ( n = 5 mice/group, p = 0.0219). d At left, representative flow cytometric analysis of CD45 − VE-cad + BM ECs within BM lin − cells from control mice and at day +7 following 5-FU chemotherapy and anti-NRP1 or IgG treatment. At right, percentages and numbers of BM ECs at day +7 following 5-FU chemotherapy and the treatments shown. Dotted lines represent numbers in control mice ( n = 5 mice/group; % BM ECs p < .0001; number BM ECs p = 0.0008). e At left, representative histograms show % activated caspase 3/7 + cells within lin − CD45 − VE-cad + BM ECs at day +7 following 5-FU. At right, % activated caspase 3/7 + BM ECs at day +7 ( n = 5/group; p < .0001). Dotted line represents percentages of activated caspase 3/7 + cells within lin − CD45 − VE-cad + BM ECs in non-IRR controls. f PB WBC, NEU, and LYMPHs at day +7 following 5-FU and the treatments shown ( n = 5 mice/group; p < 0.0001). g At left, flow cytometry of BM KSL cells at day +7 following 5-FU and the treatments shown. At right, % BM KSL cells ( n = 5 mice/group; p = 0.011). h Colony forming unit quantification from 30,000 BM cells isolated from mice at day +7 following 5-FU and IgG or anti-NRP1 treatment ( n = 4 replicates, mean values +/− SEM; GM: p < 0.0001; GEMM: p = 0.0201). Statistics show GM and GEMM comparisons. a , f , h Two-way ANOVA followed by Holm-Sidak’s multiple comparison two-sided t -tests; c , d , g Student’s unpaired two-sided t -test. * p < 0.05, *** p < 0.001, **** p < 0.0001. Source data provided as a Source Data file.

Journal: Nature Communications

Article Title: Neuropilin 1 regulates bone marrow vascular regeneration and hematopoietic reconstitution

doi: 10.1038/s41467-021-27263-y

Figure Lengend Snippet: a qRT-PCR analysis of Sema3a and Nrp1 expression in BM ECs from mice at +24 h following 5-FU chemotherapy. For each gene, expression is normalized to vehicle treatment ( n = 5/condition; Sema3a : p < 0.0001, Nrp1 : p = 0.1291). b Representative images of VE-cadherin + (magenta) , CD31 + (green) BM vessels in femur sections from untreated control mice and at day +7 following 5-FU chemotherapy and treatment with anti-NRP1 or IgG. Nuclei are stained with DAPI (blue). Scale bar, 50 µm; magnified view scale bar, 20 µm. c BM cell counts at day +7 following 5-FU chemotherapy and treatment with anti-NRP1 or IgG ( n = 5 mice/group, p = 0.0219). d At left, representative flow cytometric analysis of CD45 − VE-cad + BM ECs within BM lin − cells from control mice and at day +7 following 5-FU chemotherapy and anti-NRP1 or IgG treatment. At right, percentages and numbers of BM ECs at day +7 following 5-FU chemotherapy and the treatments shown. Dotted lines represent numbers in control mice ( n = 5 mice/group; % BM ECs p < .0001; number BM ECs p = 0.0008). e At left, representative histograms show % activated caspase 3/7 + cells within lin − CD45 − VE-cad + BM ECs at day +7 following 5-FU. At right, % activated caspase 3/7 + BM ECs at day +7 ( n = 5/group; p < .0001). Dotted line represents percentages of activated caspase 3/7 + cells within lin − CD45 − VE-cad + BM ECs in non-IRR controls. f PB WBC, NEU, and LYMPHs at day +7 following 5-FU and the treatments shown ( n = 5 mice/group; p < 0.0001). g At left, flow cytometry of BM KSL cells at day +7 following 5-FU and the treatments shown. At right, % BM KSL cells ( n = 5 mice/group; p = 0.011). h Colony forming unit quantification from 30,000 BM cells isolated from mice at day +7 following 5-FU and IgG or anti-NRP1 treatment ( n = 4 replicates, mean values +/− SEM; GM: p < 0.0001; GEMM: p = 0.0201). Statistics show GM and GEMM comparisons. a , f , h Two-way ANOVA followed by Holm-Sidak’s multiple comparison two-sided t -tests; c , d , g Student’s unpaired two-sided t -test. * p < 0.05, *** p < 0.001, **** p < 0.0001. Source data provided as a Source Data file.

Article Snippet: For Leptin receptor+(LepR + ) stromal cells, CD45 − LepR + cells (1:200, rabbit anti-PE leptin receptor antibody, bs-0961R-PE, BiosS, Woburn, MA) were isolated.

Techniques: Quantitative RT-PCR, Expressing, Staining, Flow Cytometry, Isolation

a qRT-PCR analysis for Nrp1 expression in VE-cadherin + BM ECs from Nrp1 fl/fl mice and Cdh5-Cre-ERT2;Nrp1 fl/fl mice following tamoxifen treatment ( n = 4–5 mice/group, p < 0.0001). Nrp1 expression was normalized to Gapdh levels and then replicates were normalized relative to the Nrp1 fl/fl group. b Representative microscopic images of VE-cad + BM vessels (red) in femur sections from Nrp1 fl/fl mice and Cdh5-Cre-ERT2;Nrp1 fl/fl mice at day +7 following 500 cGy TBI. Scale bar, 100 µm. c BM cell counts in Nrp1 fl/fl mice and Cdh5-Cre-ERT2;Nrp1 fl/fl mice at day +10 following 500 cGy ( n = 10 mice/group; p = 0.001). d Representative flow cytometric analysis of CD45 − CD31 + BM ECs within BM lin − cells from Nrp1 fl/fl mice and Cdh5-Cre-ERT2;Nrp1 fl/fl mice are shown at day +10. At right, representative percentages of Annexin + 7AAD − and Annexin + 7AAD + cells within the BM EC population are shown. e At left, mean percentages of CD31 + BM ECs within the BM lin − population at day +10 and at right, numbers of femoral CD31 + BM ECs at day +10 following 500 cGy TBI ( n = 5 mice/group; %BM ECs: p < 0.0001; number BM ECs: p = 0.0056). f Mean percentages of live, apoptotic, and necrotic BM ECs at day +10 following 500 cGy in the groups shown ( n = 5 mice/group; p < 0.0001). g PB WBC, NEU, and LYMPHS at day + 10 following 500 cGy TBI in Nrp1 fl/fl mice and Cdh5-Cre-ERT2;Nrp1 fl/fl mice ( n = 10 mice/group; WBC: p < 0.0001; NEU: p = 0.0014; LYMPH: p = 0.0327). h At left, representative flow cytometric analysis of BM c-kit + sca1 − lin − progenitors and KSL stem/progenitor cells at day +10 following 500 cGy TBI. At right, number of c-kit + sca1 − lin − progenitors and KSL cells at day +10 in the groups shown ( n = 10 mice/group, p < 0.0001). i CFC quantification from 10,000 BM cells isolated from mice at day +10 following IR ( n = 10 replicates, data are presented as mean values +/− SEM, p < 0.0001). a , c , e , h Student’s unpaired two-sided t -test, f , g , i Two-way ANOVA with Holm-Sidak’s multiple comparison two-sided t -tests. * p < 0.05, ** p < 0.01, **** p < 0.0001. Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Neuropilin 1 regulates bone marrow vascular regeneration and hematopoietic reconstitution

doi: 10.1038/s41467-021-27263-y

Figure Lengend Snippet: a qRT-PCR analysis for Nrp1 expression in VE-cadherin + BM ECs from Nrp1 fl/fl mice and Cdh5-Cre-ERT2;Nrp1 fl/fl mice following tamoxifen treatment ( n = 4–5 mice/group, p < 0.0001). Nrp1 expression was normalized to Gapdh levels and then replicates were normalized relative to the Nrp1 fl/fl group. b Representative microscopic images of VE-cad + BM vessels (red) in femur sections from Nrp1 fl/fl mice and Cdh5-Cre-ERT2;Nrp1 fl/fl mice at day +7 following 500 cGy TBI. Scale bar, 100 µm. c BM cell counts in Nrp1 fl/fl mice and Cdh5-Cre-ERT2;Nrp1 fl/fl mice at day +10 following 500 cGy ( n = 10 mice/group; p = 0.001). d Representative flow cytometric analysis of CD45 − CD31 + BM ECs within BM lin − cells from Nrp1 fl/fl mice and Cdh5-Cre-ERT2;Nrp1 fl/fl mice are shown at day +10. At right, representative percentages of Annexin + 7AAD − and Annexin + 7AAD + cells within the BM EC population are shown. e At left, mean percentages of CD31 + BM ECs within the BM lin − population at day +10 and at right, numbers of femoral CD31 + BM ECs at day +10 following 500 cGy TBI ( n = 5 mice/group; %BM ECs: p < 0.0001; number BM ECs: p = 0.0056). f Mean percentages of live, apoptotic, and necrotic BM ECs at day +10 following 500 cGy in the groups shown ( n = 5 mice/group; p < 0.0001). g PB WBC, NEU, and LYMPHS at day + 10 following 500 cGy TBI in Nrp1 fl/fl mice and Cdh5-Cre-ERT2;Nrp1 fl/fl mice ( n = 10 mice/group; WBC: p < 0.0001; NEU: p = 0.0014; LYMPH: p = 0.0327). h At left, representative flow cytometric analysis of BM c-kit + sca1 − lin − progenitors and KSL stem/progenitor cells at day +10 following 500 cGy TBI. At right, number of c-kit + sca1 − lin − progenitors and KSL cells at day +10 in the groups shown ( n = 10 mice/group, p < 0.0001). i CFC quantification from 10,000 BM cells isolated from mice at day +10 following IR ( n = 10 replicates, data are presented as mean values +/− SEM, p < 0.0001). a , c , e , h Student’s unpaired two-sided t -test, f , g , i Two-way ANOVA with Holm-Sidak’s multiple comparison two-sided t -tests. * p < 0.05, ** p < 0.01, **** p < 0.0001. Source data are provided as a Source Data file.

Article Snippet: For Leptin receptor+(LepR + ) stromal cells, CD45 − LepR + cells (1:200, rabbit anti-PE leptin receptor antibody, bs-0961R-PE, BiosS, Woburn, MA) were isolated.

Techniques: Quantitative RT-PCR, Expressing, Isolation

a (Left) flow cytometry of activated caspase 3/7 + BM ECs in wild-type (WT) and Nrp1 VEGF− mice at day +5 following 500 cGy and the treatments shown; (Right) % activated caspase 3/7 + BM ECs ( n = 5/group; **** p < 0.0001; *** p = 0.0003). b (Left) Donor CD45.2 + cell engraftment at 20 weeks in the PB of CD45.1 + mice competitively transplanted with BM cells from 500 cGy—irradiated Nrp1 VEGF− mice treated with anti-NRP1 or IgG; (Right) % donor CD45.2 + , Mac1 + /Gr1 + , B220 + , and CD3 + cells at 20 weeks ( n = 5–6/group; total: p = 0.0346; myeloid: p = 0.0325; T cells: p = 0.0195). c (Left) Histograms of p-Cdk5 + BM ECs at 1 h following 800 cGy and the treatments shown; (Right) %p-p53 + BM ECs. Horizontal lines show gating for p-Cdk5 + cells ( n = 5 replicates, Non-IRR; n = 6 replicates for IRR, IRR + SEMA3A, IRR + SEMA3A + anti-NRP1; non-IRR vs. IRR: p < 0.0001; IRR vs. IRR + SEMA3A: p = 0.0005; IRR vs. IRR + SEMA3A + Anti-NRP1 p < 0.0001). d % activated caspase 3/7 + BM ECs at 24 h following 800 cGy and treatment with SEMA3A, SEMA3A + anti-NRP1 or SEMA3A + roscovitine (ROSC), 10 ng/ml ( n = 4–8 replicates; non-IRR vs. IRR: p < 0.0001; IRR vs. IRR + SEMA3A: p = 0.0007; IRR vs. IRR + SEMA3A + Anti-NRP1 p < 0.0001; IRR vs. IRR + SEMA3A + Anti-NRP1 + ROSC p < 0.0001). e At left, histograms of p-p53 + BM ECs at 1 h following 800 cGy and treatment with SEMA3A, SEMA3A + anti-NRP1, or SEMA3A + ROSC. At right, % p-p53 + BM ECs are shown. Black horizontal lines show gating for p-p53 + cells ( n = 6 replicates, Non-IRR; n = 9 replicates for IRR and IRR + SEMA3A; n = 5 replicates for IRR + SEMA3A + anti-NRP1; n = 4 replicates for IRR + SEMA3A + ROSC; p < 0.0001 for all comparisons). f At left, flow cytometry for activated caspase 3/7 + BM ECs from p53 −/− and p53 + /+ mice at 24 h following irradiation with 800 cGy and the treatments shown. At right, % activated caspase 3/7 + BM ECs are shown ( n = 5 replicates/group, p < 0.0001). g Puma expression in BM ECs at 6 h following 800 cGy and the treatments shown ( n = 4/condition; **** p < 0.0001, * p = 0.0142, ** p = 0.0025, *** p = 0.0009). a , c , d , e One-way ANOVA followed by Holm-Sidak’s two-sided unpaired t -tests, f Two-way ANOVA followed by Holm-Sidak’s two-sided unpaired t -test, g Brown–Forsythe ANOVA followed by two-sided unpaired t -tests with Welch’s correction, b Two-sided Mann–Whitney test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Source Data file provided.

Journal: Nature Communications

Article Title: Neuropilin 1 regulates bone marrow vascular regeneration and hematopoietic reconstitution

doi: 10.1038/s41467-021-27263-y

Figure Lengend Snippet: a (Left) flow cytometry of activated caspase 3/7 + BM ECs in wild-type (WT) and Nrp1 VEGF− mice at day +5 following 500 cGy and the treatments shown; (Right) % activated caspase 3/7 + BM ECs ( n = 5/group; **** p < 0.0001; *** p = 0.0003). b (Left) Donor CD45.2 + cell engraftment at 20 weeks in the PB of CD45.1 + mice competitively transplanted with BM cells from 500 cGy—irradiated Nrp1 VEGF− mice treated with anti-NRP1 or IgG; (Right) % donor CD45.2 + , Mac1 + /Gr1 + , B220 + , and CD3 + cells at 20 weeks ( n = 5–6/group; total: p = 0.0346; myeloid: p = 0.0325; T cells: p = 0.0195). c (Left) Histograms of p-Cdk5 + BM ECs at 1 h following 800 cGy and the treatments shown; (Right) %p-p53 + BM ECs. Horizontal lines show gating for p-Cdk5 + cells ( n = 5 replicates, Non-IRR; n = 6 replicates for IRR, IRR + SEMA3A, IRR + SEMA3A + anti-NRP1; non-IRR vs. IRR: p < 0.0001; IRR vs. IRR + SEMA3A: p = 0.0005; IRR vs. IRR + SEMA3A + Anti-NRP1 p < 0.0001). d % activated caspase 3/7 + BM ECs at 24 h following 800 cGy and treatment with SEMA3A, SEMA3A + anti-NRP1 or SEMA3A + roscovitine (ROSC), 10 ng/ml ( n = 4–8 replicates; non-IRR vs. IRR: p < 0.0001; IRR vs. IRR + SEMA3A: p = 0.0007; IRR vs. IRR + SEMA3A + Anti-NRP1 p < 0.0001; IRR vs. IRR + SEMA3A + Anti-NRP1 + ROSC p < 0.0001). e At left, histograms of p-p53 + BM ECs at 1 h following 800 cGy and treatment with SEMA3A, SEMA3A + anti-NRP1, or SEMA3A + ROSC. At right, % p-p53 + BM ECs are shown. Black horizontal lines show gating for p-p53 + cells ( n = 6 replicates, Non-IRR; n = 9 replicates for IRR and IRR + SEMA3A; n = 5 replicates for IRR + SEMA3A + anti-NRP1; n = 4 replicates for IRR + SEMA3A + ROSC; p < 0.0001 for all comparisons). f At left, flow cytometry for activated caspase 3/7 + BM ECs from p53 −/− and p53 + /+ mice at 24 h following irradiation with 800 cGy and the treatments shown. At right, % activated caspase 3/7 + BM ECs are shown ( n = 5 replicates/group, p < 0.0001). g Puma expression in BM ECs at 6 h following 800 cGy and the treatments shown ( n = 4/condition; **** p < 0.0001, * p = 0.0142, ** p = 0.0025, *** p = 0.0009). a , c , d , e One-way ANOVA followed by Holm-Sidak’s two-sided unpaired t -tests, f Two-way ANOVA followed by Holm-Sidak’s two-sided unpaired t -test, g Brown–Forsythe ANOVA followed by two-sided unpaired t -tests with Welch’s correction, b Two-sided Mann–Whitney test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Source Data file provided.

Article Snippet: For Leptin receptor+(LepR + ) stromal cells, CD45 − LepR + cells (1:200, rabbit anti-PE leptin receptor antibody, bs-0961R-PE, BiosS, Woburn, MA) were isolated.

Techniques: Flow Cytometry, Irradiation, Expressing, MANN-WHITNEY

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