Journal: Cell stem cell

Article Title: Distinct Bone Marrow Sources of Pleiotrophin Control Hematopoietic Stem Cell Maintenance and Regeneration

doi: 10.1016/j.stem.2018.07.003

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit Anti-Leptin receptor Polyclonal Antibody, FITC , Bioss Antibodies , Cat. bs-0961R-FITC; RRID:AB_11074292.

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

Journal: Frontiers in Immunology

Article Title: Leptin deficiency in CD8 + T cells ameliorates non-segmental vitiligo by reducing interferon-γ and Granzyme B

doi: 10.3389/fimmu.2023.1158883

Figure Lengend Snippet: LEPR expression in CD8 + T cells from vitiligo patients was increased. (A, B) Double immunofluorescence staining results of LEPR and CD8 in vitiligo lesions and normal skin (Vitiligo: N = 5, NC: N = 5). (A) CD8 was represented by green fluorescence and LEPR was represented by red fluorescence. (B) The expression of LEPR in CD8 + T cells in Vitiligo lesions was significantly higher. (C) The serum levels of leptin were evaluated by ELISA (Vitiligo: N=53, HC: N=36). (D, E) Representative flow cytometric profiles and data plots of CD8 + T cells in PBMCs are shown (Vitiligo: N=5, HC: N=5). * P < 0.05, ** P < 0.01.

Article Snippet: The following antibodies were used: Alexa Flour 647 Donkey anti-rabbit IgG (BioLegend), Rabbit Anti-Leptin receptor antibody (bs-0410R, Bioss), APC-Cy7 anti-human CD4 (BD Biosciences), Percp-Cy5.5 anti-human CD8 (BD Biosciences), PE anti-human Perforin (BD Biosciences), FITC anti-human/mouse Granzyme B (Granzyme B, BioLegend), PE-Cy7 anti-human IFN-γ (BioLegend), Zombie Aqua Fixable Viability Kit (BioLegend), APC-Cy7 anti-mouse CD4 (Abcam), Percp-Cy5.5 anti-mouse CD8 (Abcam), PE anti-mouse Perforin (Abcam), FITC anti-mouse/human granzyme B (BioLegend), PE-Cy7 anti-mouse IFN-γ (Abcam).

Techniques: Expressing, Double Immunofluorescence Staining, Fluorescence, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Leptin deficiency in CD8 + T cells ameliorates non-segmental vitiligo by reducing interferon-γ and Granzyme B

doi: 10.3389/fimmu.2023.1158883

Figure Lengend Snippet: Leptin enhances the expression of cytotoxic cytokines from CD8 + T cells in vitro . (A, B) Gating strategy, representative flow cytometric plots, and statistical analysis of the percentage of cell subsets were shown. (C) After 72 hours of leptin stimulation, the protein levels of IFN-γ, perforin, and Granzyme B from normal PBMCs were detected by ELISA. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The following antibodies were used: Alexa Flour 647 Donkey anti-rabbit IgG (BioLegend), Rabbit Anti-Leptin receptor antibody (bs-0410R, Bioss), APC-Cy7 anti-human CD4 (BD Biosciences), Percp-Cy5.5 anti-human CD8 (BD Biosciences), PE anti-human Perforin (BD Biosciences), FITC anti-human/mouse Granzyme B (Granzyme B, BioLegend), PE-Cy7 anti-human IFN-γ (BioLegend), Zombie Aqua Fixable Viability Kit (BioLegend), APC-Cy7 anti-mouse CD4 (Abcam), Percp-Cy5.5 anti-mouse CD8 (Abcam), PE anti-mouse Perforin (Abcam), FITC anti-mouse/human granzyme B (BioLegend), PE-Cy7 anti-mouse IFN-γ (Abcam).

Techniques: Expressing, In Vitro, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Leptin deficiency in CD8 + T cells ameliorates non-segmental vitiligo by reducing interferon-γ and Granzyme B

doi: 10.3389/fimmu.2023.1158883

Figure Lengend Snippet: Leptin deficiency in CD8 + T cells ameliorated vitiligo development in mice. (A) The hair decolorization areas of mice with vitiligo induced by monobenzone are shown, marked with red circles. (B) HE staining showed that the inflammatory cells of the Lep KO mice were significantly reduced compared with the WT group. (C) The heatmap shows the differential expression of vitiligo-related genes between WT and Lep KO mice after monobenzone was administrated. (D) RT-qPCR was used to verify the differentially expressed genes in CD8 + T cells from spleens of vitiligo mice induced by monobenzone in the Lep KO group and WT group. * P < 0.05, *** P < 0.001.

Article Snippet: The following antibodies were used: Alexa Flour 647 Donkey anti-rabbit IgG (BioLegend), Rabbit Anti-Leptin receptor antibody (bs-0410R, Bioss), APC-Cy7 anti-human CD4 (BD Biosciences), Percp-Cy5.5 anti-human CD8 (BD Biosciences), PE anti-human Perforin (BD Biosciences), FITC anti-human/mouse Granzyme B (Granzyme B, BioLegend), PE-Cy7 anti-human IFN-γ (BioLegend), Zombie Aqua Fixable Viability Kit (BioLegend), APC-Cy7 anti-mouse CD4 (Abcam), Percp-Cy5.5 anti-mouse CD8 (Abcam), PE anti-mouse Perforin (Abcam), FITC anti-mouse/human granzyme B (BioLegend), PE-Cy7 anti-mouse IFN-γ (Abcam).

Techniques: Staining, Expressing, Quantitative RT-PCR

Journal: Pathology, research and practice

Article Title: Clinical - pathological significance of leptin receptor (LEPR) expression in squamous cell carcinoma of the skin.

doi: 10.1016/j.prp.2020.153111

Figure Lengend Snippet: Fig. 1. Representative images for cSCC, adjacent non-tumor tissue, LEPR expression and Ki67 expression: a) and d) absence of LEPR expression in tumor and adjacent non-tumor tissue, b) and e) moderate LEPR expression in tumor and adjacent non-tumor tissue, c) pronounced LEPR expression in tumor tissue; f) and i) low Ki67 expression index in tumor and adjacent non-tumor tissue, d) high Ki67 expression index in tumor tissue, h) absence of Ki67 expression in adjacent non-tumor tissue; (ABC x 200).

Article Snippet: Antigen retrieval was enhanced by autoclaving slides in sodium citrate buffer (pH 6.0) for 30 min. Endogenous peroxidase activity was blocked by 0.3 % hydrogen peroxide-methanol buffer for 25 min. Rabbit polyclonal anti-Leptin receptor antibody (1:60, Abcam, Burlingame CA, USA) and rabbit monoclonal Ki67 antibody (1:100, Abcam, Burlingame CA, USA) were incubated at +4 °C overnight.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Gender Dimorphism in Skeletal Muscle Leptin Receptors, Serum Leptin and Insulin Sensitivity

doi: 10.1371/journal.pone.0003466

Figure Lengend Snippet: Body composition, basal plasma glucose and endocrine variables.

Article Snippet: The polyclonal rabbit anti-human leptin receptor antibody that recognizes the human leptin receptor was obtained from Linco Research (St. Charles, Missouri, USA).

Techniques: Clinical Proteomics

Journal: PLoS ONE

Article Title: Gender Dimorphism in Skeletal Muscle Leptin Receptors, Serum Leptin and Insulin Sensitivity

doi: 10.1371/journal.pone.0003466

Figure Lengend Snippet: Relationship between the basal leptin concentration in serum and body mass index (BMI), whole body fat mass, and percentage of body fat.

Article Snippet: The polyclonal rabbit anti-human leptin receptor antibody that recognizes the human leptin receptor was obtained from Linco Research (St. Charles, Missouri, USA).

Techniques: Concentration Assay

Journal: PLoS ONE

Article Title: Gender Dimorphism in Skeletal Muscle Leptin Receptors, Serum Leptin and Insulin Sensitivity

doi: 10.1371/journal.pone.0003466

Figure Lengend Snippet: Total Protein extracts were prepared from male and female muscle and OB-R, perilipin A and alpha-tubulin protein expression was analyzed by Western blot. A: a representative immunoblot assay after incubation with a polyclonal rabbit anti-OB-R antibody specifically raised against de long isoform of the leptin receptor. B: a representative western blot after incubation with a polyclonal rabbit anti-perilipin A antibody in the same samples used in A. C: a representative immunoblot analysis after incubation with the monoclonal mouse anti-alpha-tubulin antibody in the same samples used in A. D: densitometric immunosignal values (arbitrary units of band densities) of OB-R bands relative to those obtained for alpha-tubulin.

Article Snippet: The polyclonal rabbit anti-human leptin receptor antibody that recognizes the human leptin receptor was obtained from Linco Research (St. Charles, Missouri, USA).

Techniques: Expressing, Western Blot, Incubation

Journal: PLoS ONE

Article Title: Dietary protein restriction increases hepatic leptin receptor mRNA and plasma soluble leptin receptor in male rodents

doi: 10.1371/journal.pone.0219603

Figure Lengend Snippet: Primers for realtime PCR.

Article Snippet: Anti-leptin receptor polyclonal antibody (Novus Biologicals, NB120-5593, 1:2000 dilution) and anti-β-actin monoclonal antibody (Santa Cruz Biotechnology, sc-69879, 1:500 dilution) were used as 1 st antibodies and goat anti-rabbit IgG-HRP (Santa Cruz, sc-2004, 1:50000 dilution) and goat anti-mouse IgG-HRP (Santa Cruz, sc-2005, 1:50000 dilution) were used as 2 nd antibodies, respectively.

Techniques: Sequencing

Journal: PLoS ONE

Article Title: Dietary protein restriction increases hepatic leptin receptor mRNA and plasma soluble leptin receptor in male rodents

doi: 10.1371/journal.pone.0219603

Figure Lengend Snippet: (A) Rats were fed a control diet (20C) or a low-protein diet (5C) for 7 d and sacrificed after 12-h fasting (20CF, 5CF) or 12-h fasting followed by 12-h re-feeding (20CR, 5CR). Leptin receptor mRNA was measured by real-time PCR and results were expressed relative to that of 20CF, means ± standard errors (n = 5). Results of two-way ANOVA are given below the graph (NS, not significant; **, p < 0.01). (B) Rats were fed a control diet ad libitum (20C), fed a low-protein diet ad libitum (5C), or pair-fed a control diet with 5C (20R) for 16 d. Ob-R mRNA was measured by real-time PCR and expressed as means ± standard errors (n = 5). Results with different alphabet are statistically different ( P < 0.05).

Article Snippet: Anti-leptin receptor polyclonal antibody (Novus Biologicals, NB120-5593, 1:2000 dilution) and anti-β-actin monoclonal antibody (Santa Cruz Biotechnology, sc-69879, 1:500 dilution) were used as 1 st antibodies and goat anti-rabbit IgG-HRP (Santa Cruz, sc-2004, 1:50000 dilution) and goat anti-mouse IgG-HRP (Santa Cruz, sc-2005, 1:50000 dilution) were used as 2 nd antibodies, respectively.

Techniques: Control, Real-time Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: Dietary protein restriction increases hepatic leptin receptor mRNA and plasma soluble leptin receptor in male rodents

doi: 10.1371/journal.pone.0219603

Figure Lengend Snippet: C57BL/6 mice were fed 20C or 5C ad libitum for 7 d. Plasma sOb-R (A) and plasma leptin (B) concentrations were measured with ELISA. Free leptin index was calculated from plasma leptin and sOb-R (C). Results are expressed as means ± standard errors (n = 5). Statistical difference between the groups are shown as **, P < 0.01.

Article Snippet: Anti-leptin receptor polyclonal antibody (Novus Biologicals, NB120-5593, 1:2000 dilution) and anti-β-actin monoclonal antibody (Santa Cruz Biotechnology, sc-69879, 1:500 dilution) were used as 1 st antibodies and goat anti-rabbit IgG-HRP (Santa Cruz, sc-2004, 1:50000 dilution) and goat anti-mouse IgG-HRP (Santa Cruz, sc-2005, 1:50000 dilution) were used as 2 nd antibodies, respectively.

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay

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

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

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

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

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

Journal: Reproductive Sciences

Article Title: Female Infertility and Risk for Later-Life Cardiovascular Disease: Lessons from a Mouse Model of Human Cardiovascular Disease

doi: 10.1007/s43032-025-02026-y

Figure Lengend Snippet: Immunohistochemical evaluation of uteri on day 4.5 post-coitus in SR-BI KO/ ApoeR61 h/h mice. Leptin receptor (Leptin R) ( a-c ), cyclooxygenase (COX) −2 ( d-f ), leukaemia inhibitory factor (LIF) ( g-i ) and phospho-signal transducer and activator of transcription (Stat) −3 ( j-l ) immunostaining with haematoxylin counterstaining in SR-BI KO/ ApoeR61 h/h mice with placebo ( b , e , h , k ) or probucol treatment ( c , f , i , l ). As a control, uteri from wild type mice ( a , d , g , j ) were evaluated. Scale bar = 100 µm

Article Snippet: The 5-μm paraffin-embedded tissue sections were immunostained using monoclonal rabbit anti-mouse phosphorylated signal transducer and activator of transcription-3 (p-Stat3) antibody (Tyr705) (#9145; Cell Signaling Technology, Tokyo, Japan) at 1:400 dilution, polyclonal rabbit anti-mouse leukaemia inhibitory factor (LIF) antibody (OABF00432; Aviva Systems biology, CA, US) at 1:400 dilution, polyclonal goat anti-mouse leptin receptor antibody (AF497; R&D Systems, MN, US) at 1:200 dilution, and polyclonal rabbit anti-mouse cyclooxygenase (COX) −2 antibody (#160,126; Cayman Chemical, MI, US) at 1:500 dilution according to the manufacturers’ instructions.

Techniques: Immunohistochemical staining, Immunostaining, Control