anti-sca-1 Search Results


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  • 91
    Miltenyi Biotec anti sca 1
    Immunophenotyping and survival of group 2 innate lymphoid cell (ILC2)-deficient mice after cecal ligation and puncture (CLP) surgery. A: Flow cytometric data showing the elimination of lineage − /propidium iodide (PI) − CD127 + CD25 + CD117 − cells that are double positive for <t>Sca-1</t> and ST2 in the lamina propria (LP) of the small intestine (s. int.) after CLP surgery. B: Survival study demonstrating a persistent survival advantage for mice deficient in ILC2s after CLP surgery. C: Bacterial burden assay showing a significantly lower bacterial count in the peritoneal fluid after CLP surgery in ILC2-deficient mice than in control mice, but no significant difference was found in the whole blood. D: Changes in tumor necrosis factor (TNF)-α, IL-6, IL-1β, and IL-10 levels were shown in the peritoneal fluid after CLP surgery in control (CB6F1/J mice) and ILC2-deficient mice. Data are expressed as means ± SEM. n = 19 to 20 mice per group ( B ); n = 3 to 5 mice per group ( C ); n = 6 to 8 mice per group ( D ). ∗ P
    Anti Sca 1, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 91/100, based on 48 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    BioLegend anti sca 1
    Phenotype of hepatic ILC2 in homeostasis and after IL-33 treatment. C57BL/6 mice were treated with rmIL-33 on four consecutive days. ( a ) Hepatic leukocytes from naive and IL-33-treated mice were stained for lin − <t>Sca-1</t> + ST2 + ILC2 and analysed by flow cytometry. Dot plots show frequencies of ILC2 in the liver. ( b ) Hepatic ILC2 were stained for KLRG1, CD25, and GATA3 and analysed by flow cytometry. ( c ) Hepatic mRNA expression was analysed by quantitative RT-PCR in relation to the reference gene GAPDH. ( d ) Hepatic ILC2 were isolated from IL-33-treated mice by FACS. Cytokine mRNA and protein expression were analysed by quantitative RT-PCR in relation to the reference gene GAPDH and by flow cytometry, respectively. Histograms show frequencies of protein-expressing hepatic ILC2. Bold line, antibody staining; filled graph, fluorescence minus one control. Mean ± SEM of 3–4 independent experiments with 3 mice per group are shown. Mann-Whitney U test. *p
    Anti Sca 1, supplied by BioLegend, used in various techniques. Bioz Stars score: 91/100, based on 141 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Becton Dickinson anti sca 1
    In vitro expansion and isolation of Lin − <t>Sca-1</t> + c-Kit + HSPCs. ( a ) In vitro expansion of HSCs. Bone marrow-derived HSPCs were expanded by treatment with FLT3L (100 ngml −1 ), TPO (100 ngml −1 ) and SCF (100 ngml −1 ) for 27 d. The expanded cell populations were analyzed via flow cytometry with anti-Lin, Sca-1 and c-Kit antibodies (left panel). Lin − Sca-1 + c-Kit + (LSK) cells were isolated from 27 d of expanded culture using an HSC isolation kit. The isolated LSK cells were analyzed via flow cytometry with anti-Lin, Sca-1 and c-Kit antibodies (right panel). ( b ) The contamination of isolated LSK cells by B-lineage cells. Isolated Lin − Sca-1 + c-Kit + HSPCs were analyzed via flow cytometry with anti-B220, CD19, CD43, CD2, IgM, IgD and CD93 antibodies. ( c ) Analysis of isolated Lin − Sca-1 + c-Kit + HSPCs. Lin − gated cells (bolded box) were analyzed via flow cytometry with anti-Sca-1 and c-Kit antibodies (left top panel). For the analysis of the HSC, MPP and LMPP populations of HSPCs, the LSK cells were analyzed with anti-c-Kit and Flk2 antibodies (left middle panel). The averaged percentages of the HSC, MPP and LMPP populations among HSPCs were summarized (right top panel). The populations of LT-HSC and ST-HSC in the HSC (c-Kit + Flk2 − )-gated cells (bolded box) were further analyzed with anti-CD34 antibody (left bottom panel). The averaged percentages of LT-HSC and ST-HSC population in HSC-gated cells were summarized (right bottom panel). ** P
    Anti Sca 1, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 91/100, based on 286 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Thermo Fisher anti sca 1
    The change in the spatial distribution of H4K16ac upon aging is linked to a change in HSC function. a ]. b Representative FACS dot plots of HSCs <t>(Lin-c-kit+Sca-1+Flk2-CD34-),</t> ST-HSCs (Lin-c-kit+Sca-1+Flk2-CD34+), LMPPs (Lin-c-kit+Sca-1+Flk2+CD34+), and LSKs (Lin-c-kit+Sca-1+) gating strategy of young and aged lineage-depleted BM cells. c Percentage of young and aged HSCs with a polar distribution of H4K16ac, H4K8ac, H4K5ac, H3K27ac, H3K4me1, and H3K4me3; n = 3–4 biological repeats; ~ 150–200 single HSCs scored per sample in total; * p
    Anti Sca 1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 396 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Santa Cruz Biotechnology anti sca 1
    The change in the spatial distribution of H4K16ac upon aging is linked to a change in HSC function. a ]. b Representative FACS dot plots of HSCs <t>(Lin-c-kit+Sca-1+Flk2-CD34-),</t> ST-HSCs (Lin-c-kit+Sca-1+Flk2-CD34+), LMPPs (Lin-c-kit+Sca-1+Flk2+CD34+), and LSKs (Lin-c-kit+Sca-1+) gating strategy of young and aged lineage-depleted BM cells. c Percentage of young and aged HSCs with a polar distribution of H4K16ac, H4K8ac, H4K5ac, H3K27ac, H3K4me1, and H3K4me3; n = 3–4 biological repeats; ~ 150–200 single HSCs scored per sample in total; * p
    Anti Sca 1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Abcam anti sca 1
    Characterization of MSC mix and NCSC mix isolated from the bone marrow of adult Wnt1-CRE/R26R-LacZ mice. After recombination, NCSCs from Wnt1-CRE/R26R-LacZ mice express LacZ gene. MSCs did not undergo Cre/Lox recombination and conserved the PGK-Neo cassette ( a ). MSC mix are adherent fibroblast-like cells, do not express β-galactosidase ( b ) or Sox2 c (red), slightly express Nestin ( c ) (green), p75NTR ( d ) (red), and <t>Sca-1</t> ( d ) (green). NCSC mix have a similar morphology, express β-galactosidase ( e ), Nestin ( f ) (green), Sox2 ( f ) (red) and p75NTR ( g ) red), but not Sca-1( g ) (green). Scale bar = 20 μm. MSC mesenchymal stem cell, NCSC neural crest stem cell
    Anti Sca 1, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher apc anti sca 1
    Characterization of MSC mix and NCSC mix isolated from the bone marrow of adult Wnt1-CRE/R26R-LacZ mice. After recombination, NCSCs from Wnt1-CRE/R26R-LacZ mice express LacZ gene. MSCs did not undergo Cre/Lox recombination and conserved the PGK-Neo cassette ( a ). MSC mix are adherent fibroblast-like cells, do not express β-galactosidase ( b ) or Sox2 c (red), slightly express Nestin ( c ) (green), p75NTR ( d ) (red), and <t>Sca-1</t> ( d ) (green). NCSC mix have a similar morphology, express β-galactosidase ( e ), Nestin ( f ) (green), Sox2 ( f ) (red) and p75NTR ( g ) red), but not Sca-1( g ) (green). Scale bar = 20 μm. MSC mesenchymal stem cell, NCSC neural crest stem cell
    Apc Anti Sca 1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher anti sca 1 pecy7
    Characterization of MSC mix and NCSC mix isolated from the bone marrow of adult Wnt1-CRE/R26R-LacZ mice. After recombination, NCSCs from Wnt1-CRE/R26R-LacZ mice express LacZ gene. MSCs did not undergo Cre/Lox recombination and conserved the PGK-Neo cassette ( a ). MSC mix are adherent fibroblast-like cells, do not express β-galactosidase ( b ) or Sox2 c (red), slightly express Nestin ( c ) (green), p75NTR ( d ) (red), and <t>Sca-1</t> ( d ) (green). NCSC mix have a similar morphology, express β-galactosidase ( e ), Nestin ( f ) (green), Sox2 ( f ) (red) and p75NTR ( g ) red), but not Sca-1( g ) (green). Scale bar = 20 μm. MSC mesenchymal stem cell, NCSC neural crest stem cell
    Anti Sca 1 Pecy7, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Pharmingen anti sca 1 pe
    hCMPC in the human heart. (A-G) Immunohistochemistry for <t>Sca-1</t> in foetal and adult heart. (B, C) High power magnification of areas in A. (D) Atrial ventricular boundary. (E) High power magnification of area in D. (F, H) hCMPCs in biopsy from adult patient. (G) High power magnification of area in F. (I) IgG control. Arrows designate some of the hCMPCs. Magnification: A, D, F, H, I: 100x, B, C, E, G: 200x.
    Anti Sca 1 Pe, supplied by Pharmingen, used in various techniques. Bioz Stars score: 89/100, based on 54 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Boster Bio anti sca 1 antibody
    (a) Culture of muscle-derived stem cells. MDSCs were cultured and subcultured at confluent of 80%. Bright view images were taken at day 1 and day 5 after subconfluent. (b) Immunohistochemical staining of <t>Sca-1.</t> MDSCs were subcultured. Expression of Sca-1 was assessed by immunohistochemical staining analysis at day 2 using an anti-Sca-1 antibody (Wuhan Boster Biological Technology Co., Ltd.).
    Anti Sca 1 Antibody, supplied by Boster Bio, used in various techniques. Bioz Stars score: 88/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    BioLegend pecy7 anti sca 1
    (a) Culture of muscle-derived stem cells. MDSCs were cultured and subcultured at confluent of 80%. Bright view images were taken at day 1 and day 5 after subconfluent. (b) Immunohistochemical staining of <t>Sca-1.</t> MDSCs were subcultured. Expression of Sca-1 was assessed by immunohistochemical staining analysis at day 2 using an anti-Sca-1 antibody (Wuhan Boster Biological Technology Co., Ltd.).
    Pecy7 Anti Sca 1, supplied by BioLegend, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher anti sca 1 d7
    (a) Culture of muscle-derived stem cells. MDSCs were cultured and subcultured at confluent of 80%. Bright view images were taken at day 1 and day 5 after subconfluent. (b) Immunohistochemical staining of <t>Sca-1.</t> MDSCs were subcultured. Expression of Sca-1 was assessed by immunohistochemical staining analysis at day 2 using an anti-Sca-1 antibody (Wuhan Boster Biological Technology Co., Ltd.).
    Anti Sca 1 D7, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 115 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Becton Dickinson rat anti sca 1
    Prolonged expression of Sox17 results in formation of bronchiolar-like structures in the alveoli. Adult CCSPrtTA/tetO-Sox17 transgenic mice were maintained on Dox for 12 months. (A) H E staining shows the presence of bronchiolar-like sheets of cells in the peripheral lung. Arrowhead indicates the pleural surface. (B–D) Immunostaining for Sox17 (B), Foxj1 (C), and CCSP (D) was performed on lung sections. The Sox17-induced bronchiolar-like structures contained cells expressing proximal airway markers CCSP and Foxj1. (E–H) Immunofluorescent staining for CCSP (E), proSP-C (F), and <t>Sca-1</t> (G). The bronchiolar-like structures (dotted outline) contained cells that express Sca-1 and subsets of cells expressing CCSP or proSP-C. Arrowheads demark CCSP-expressing cells in the bronchiolar epithelium and the arrow indicates normal proSP-C expression in a type II cell. Nuclei are stained with DAPI (H; blue) Scale bars, 50 µm.
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    Image Search Results


    Immunophenotyping and survival of group 2 innate lymphoid cell (ILC2)-deficient mice after cecal ligation and puncture (CLP) surgery. A: Flow cytometric data showing the elimination of lineage − /propidium iodide (PI) − CD127 + CD25 + CD117 − cells that are double positive for Sca-1 and ST2 in the lamina propria (LP) of the small intestine (s. int.) after CLP surgery. B: Survival study demonstrating a persistent survival advantage for mice deficient in ILC2s after CLP surgery. C: Bacterial burden assay showing a significantly lower bacterial count in the peritoneal fluid after CLP surgery in ILC2-deficient mice than in control mice, but no significant difference was found in the whole blood. D: Changes in tumor necrosis factor (TNF)-α, IL-6, IL-1β, and IL-10 levels were shown in the peritoneal fluid after CLP surgery in control (CB6F1/J mice) and ILC2-deficient mice. Data are expressed as means ± SEM. n = 19 to 20 mice per group ( B ); n = 3 to 5 mice per group ( C ); n = 6 to 8 mice per group ( D ). ∗ P

    Journal: The American Journal of Pathology

    Article Title: Group 2 Innate Lymphoid Cells (ILC2s) Are Key Mediators of the Inflammatory Response in Polymicrobial Sepsis

    doi: 10.1016/j.ajpath.2018.05.009

    Figure Lengend Snippet: Immunophenotyping and survival of group 2 innate lymphoid cell (ILC2)-deficient mice after cecal ligation and puncture (CLP) surgery. A: Flow cytometric data showing the elimination of lineage − /propidium iodide (PI) − CD127 + CD25 + CD117 − cells that are double positive for Sca-1 and ST2 in the lamina propria (LP) of the small intestine (s. int.) after CLP surgery. B: Survival study demonstrating a persistent survival advantage for mice deficient in ILC2s after CLP surgery. C: Bacterial burden assay showing a significantly lower bacterial count in the peritoneal fluid after CLP surgery in ILC2-deficient mice than in control mice, but no significant difference was found in the whole blood. D: Changes in tumor necrosis factor (TNF)-α, IL-6, IL-1β, and IL-10 levels were shown in the peritoneal fluid after CLP surgery in control (CB6F1/J mice) and ILC2-deficient mice. Data are expressed as means ± SEM. n = 19 to 20 mice per group ( B ); n = 3 to 5 mice per group ( C ); n = 6 to 8 mice per group ( D ). ∗ P

    Article Snippet: Cells were stained with propidium iodide (PI) to exclude nonviable cells and with antilineage antibodies, including anti-CD3ε, CD19, NK1.1, Gr-1, Ter119, CD11b antibodies, anti-CD45, anti-CD127, anti-CD117, anti-CD25, anti-CCR6, anti–Sca-1 (Miltenyi Biotec Inc.), and anti-IL33Rα (ST2) (BioLegend, San Diego, CA) antibodies, as well as with the appropriate isotope control antibodies (Miltenyi Biotec Inc.).

    Techniques: Mouse Assay, Ligation, Flow Cytometry

    Changes in the number and percentage of group 2 innate lymphoid cells (ILC2s) among the lamina propria (LP) cells (LPCs) of the small intestine (s. int.), the nonparenchymal cells (NPCs) of the liver, the peritoneal (perit.) cells, and the splenocytes (Spl.). A: Flow cytometric analysis showing Lineage − /propidium iodide (PI) − CD127 + CD25 + CD117 − (LPCs and liver NPCs) and Lineage − /PI − CD127 + CD117 − (peritoneal cells and splenocytes) cells that were double positive for Sca-1 and ST2 ( boxed areas denote ILC2s) 24 hours after sham or cecal ligation and puncture (CLP) surgery. B–E: ILC2s changed in their number and proportion at different tissue/fluid compartments after CLP surgery. Data are expressed as means ± SEM. n = 7 to12 mice for all groups. ∗ P

    Journal: The American Journal of Pathology

    Article Title: Group 2 Innate Lymphoid Cells (ILC2s) Are Key Mediators of the Inflammatory Response in Polymicrobial Sepsis

    doi: 10.1016/j.ajpath.2018.05.009

    Figure Lengend Snippet: Changes in the number and percentage of group 2 innate lymphoid cells (ILC2s) among the lamina propria (LP) cells (LPCs) of the small intestine (s. int.), the nonparenchymal cells (NPCs) of the liver, the peritoneal (perit.) cells, and the splenocytes (Spl.). A: Flow cytometric analysis showing Lineage − /propidium iodide (PI) − CD127 + CD25 + CD117 − (LPCs and liver NPCs) and Lineage − /PI − CD127 + CD117 − (peritoneal cells and splenocytes) cells that were double positive for Sca-1 and ST2 ( boxed areas denote ILC2s) 24 hours after sham or cecal ligation and puncture (CLP) surgery. B–E: ILC2s changed in their number and proportion at different tissue/fluid compartments after CLP surgery. Data are expressed as means ± SEM. n = 7 to12 mice for all groups. ∗ P

    Article Snippet: Cells were stained with propidium iodide (PI) to exclude nonviable cells and with antilineage antibodies, including anti-CD3ε, CD19, NK1.1, Gr-1, Ter119, CD11b antibodies, anti-CD45, anti-CD127, anti-CD117, anti-CD25, anti-CCR6, anti–Sca-1 (Miltenyi Biotec Inc.), and anti-IL33Rα (ST2) (BioLegend, San Diego, CA) antibodies, as well as with the appropriate isotope control antibodies (Miltenyi Biotec Inc.).

    Techniques: Flow Cytometry, Ligation, Mouse Assay

    Phenotype of hepatic ILC2 in homeostasis and after IL-33 treatment. C57BL/6 mice were treated with rmIL-33 on four consecutive days. ( a ) Hepatic leukocytes from naive and IL-33-treated mice were stained for lin − Sca-1 + ST2 + ILC2 and analysed by flow cytometry. Dot plots show frequencies of ILC2 in the liver. ( b ) Hepatic ILC2 were stained for KLRG1, CD25, and GATA3 and analysed by flow cytometry. ( c ) Hepatic mRNA expression was analysed by quantitative RT-PCR in relation to the reference gene GAPDH. ( d ) Hepatic ILC2 were isolated from IL-33-treated mice by FACS. Cytokine mRNA and protein expression were analysed by quantitative RT-PCR in relation to the reference gene GAPDH and by flow cytometry, respectively. Histograms show frequencies of protein-expressing hepatic ILC2. Bold line, antibody staining; filled graph, fluorescence minus one control. Mean ± SEM of 3–4 independent experiments with 3 mice per group are shown. Mann-Whitney U test. *p

    Journal: Scientific Reports

    Article Title: Hepatic ILC2 activity is regulated by liver inflammation-induced cytokines and effector CD4+ T cells

    doi: 10.1038/s41598-020-57985-w

    Figure Lengend Snippet: Phenotype of hepatic ILC2 in homeostasis and after IL-33 treatment. C57BL/6 mice were treated with rmIL-33 on four consecutive days. ( a ) Hepatic leukocytes from naive and IL-33-treated mice were stained for lin − Sca-1 + ST2 + ILC2 and analysed by flow cytometry. Dot plots show frequencies of ILC2 in the liver. ( b ) Hepatic ILC2 were stained for KLRG1, CD25, and GATA3 and analysed by flow cytometry. ( c ) Hepatic mRNA expression was analysed by quantitative RT-PCR in relation to the reference gene GAPDH. ( d ) Hepatic ILC2 were isolated from IL-33-treated mice by FACS. Cytokine mRNA and protein expression were analysed by quantitative RT-PCR in relation to the reference gene GAPDH and by flow cytometry, respectively. Histograms show frequencies of protein-expressing hepatic ILC2. Bold line, antibody staining; filled graph, fluorescence minus one control. Mean ± SEM of 3–4 independent experiments with 3 mice per group are shown. Mann-Whitney U test. *p

    Article Snippet: Single cell suspensions were stained with the Lineage Antibody Cocktail (APC; BD Pharmingen, Heidelberg, Germany) as well as anti-Sca-1 (Pacific Blue; D7; BioLegend) and anti-ST2 (PerCP-eFluor 710; RMST2-2; ThermoFisher Scientific, Waltham, MA) antibodies.

    Techniques: Mouse Assay, Staining, Flow Cytometry, Expressing, Quantitative RT-PCR, Isolation, FACS, Fluorescence, MANN-WHITNEY

    In vitro expansion and isolation of Lin − Sca-1 + c-Kit + HSPCs. ( a ) In vitro expansion of HSCs. Bone marrow-derived HSPCs were expanded by treatment with FLT3L (100 ngml −1 ), TPO (100 ngml −1 ) and SCF (100 ngml −1 ) for 27 d. The expanded cell populations were analyzed via flow cytometry with anti-Lin, Sca-1 and c-Kit antibodies (left panel). Lin − Sca-1 + c-Kit + (LSK) cells were isolated from 27 d of expanded culture using an HSC isolation kit. The isolated LSK cells were analyzed via flow cytometry with anti-Lin, Sca-1 and c-Kit antibodies (right panel). ( b ) The contamination of isolated LSK cells by B-lineage cells. Isolated Lin − Sca-1 + c-Kit + HSPCs were analyzed via flow cytometry with anti-B220, CD19, CD43, CD2, IgM, IgD and CD93 antibodies. ( c ) Analysis of isolated Lin − Sca-1 + c-Kit + HSPCs. Lin − gated cells (bolded box) were analyzed via flow cytometry with anti-Sca-1 and c-Kit antibodies (left top panel). For the analysis of the HSC, MPP and LMPP populations of HSPCs, the LSK cells were analyzed with anti-c-Kit and Flk2 antibodies (left middle panel). The averaged percentages of the HSC, MPP and LMPP populations among HSPCs were summarized (right top panel). The populations of LT-HSC and ST-HSC in the HSC (c-Kit + Flk2 − )-gated cells (bolded box) were further analyzed with anti-CD34 antibody (left bottom panel). The averaged percentages of LT-HSC and ST-HSC population in HSC-gated cells were summarized (right bottom panel). ** P

    Journal: Experimental & Molecular Medicine

    Article Title: Generation of an osteoblast-based artificial niche that supports in vitro B lymphopoiesis

    doi: 10.1038/emm.2017.189

    Figure Lengend Snippet: In vitro expansion and isolation of Lin − Sca-1 + c-Kit + HSPCs. ( a ) In vitro expansion of HSCs. Bone marrow-derived HSPCs were expanded by treatment with FLT3L (100 ngml −1 ), TPO (100 ngml −1 ) and SCF (100 ngml −1 ) for 27 d. The expanded cell populations were analyzed via flow cytometry with anti-Lin, Sca-1 and c-Kit antibodies (left panel). Lin − Sca-1 + c-Kit + (LSK) cells were isolated from 27 d of expanded culture using an HSC isolation kit. The isolated LSK cells were analyzed via flow cytometry with anti-Lin, Sca-1 and c-Kit antibodies (right panel). ( b ) The contamination of isolated LSK cells by B-lineage cells. Isolated Lin − Sca-1 + c-Kit + HSPCs were analyzed via flow cytometry with anti-B220, CD19, CD43, CD2, IgM, IgD and CD93 antibodies. ( c ) Analysis of isolated Lin − Sca-1 + c-Kit + HSPCs. Lin − gated cells (bolded box) were analyzed via flow cytometry with anti-Sca-1 and c-Kit antibodies (left top panel). For the analysis of the HSC, MPP and LMPP populations of HSPCs, the LSK cells were analyzed with anti-c-Kit and Flk2 antibodies (left middle panel). The averaged percentages of the HSC, MPP and LMPP populations among HSPCs were summarized (right top panel). The populations of LT-HSC and ST-HSC in the HSC (c-Kit + Flk2 − )-gated cells (bolded box) were further analyzed with anti-CD34 antibody (left bottom panel). The averaged percentages of LT-HSC and ST-HSC population in HSC-gated cells were summarized (right bottom panel). ** P

    Article Snippet: Specific antibodies were purchased from the following commercial sources: anti-FLAG and anti-β-actin from Sigma-Aldrich; anti-B220, anti-Sca-1, anti-lineage cells (anti-Lin), anti-IgD and anti-IgM from BD Biosciences (San Jose, CA, USA); anti-Ki67, anti-Flk2, anti-CD2, anti-CD19, anti-CD23, anti-CD34, anti-CD43 and anti-CD93 from eBioscience (San Diego, CA, USA); and anti-receptor activator of nuclear factor κB ligand (RANKL) from Novus (Littleton, CO, USA).

    Techniques: In Vitro, Isolation, Derivative Assay, Flow Cytometry, Cytometry

    In vitro B lymphopoiesis of Lin − Sca-1 + c-Kit + HSPCs in coculture with OBN4 cells. ( a ) The increase in the number of non-adherent cells under coculture. LSK cells (10 5 cells per well of a 6-well plate) isolated as shown in Figure 4 were cocultured on monolayers of the OBN4 cells (6 × 10 4 cells per well) in the presence or absence of IL-7 (100 ngml −1 ) and SDF-1 (100 ngml −1 ) for 7 d. Black box, cytokine treatment. Open box, phosphate-buffered saline (PBS, non-cytokine treatment). ( b ) The increase in the number of B220 + cells under coculture. ( c ) In vitro B lymphopoiesis of HSPCs in coculture with OBN4 cells in the presence of IL-7 and SDF-1. The development of B-lineage cells was analyzed via flow cytometry with antibodies against B220, CD2, CD19, CD43, CD93, IgM and IgD during coculture with OBN4 for 7 d. The presented numbers indicate the percentage of each cell population. ( d ) The average numbers of B-lineage cells produced during coculture with OBN4 cells were summarized. NS, not significant. * P

    Journal: Experimental & Molecular Medicine

    Article Title: Generation of an osteoblast-based artificial niche that supports in vitro B lymphopoiesis

    doi: 10.1038/emm.2017.189

    Figure Lengend Snippet: In vitro B lymphopoiesis of Lin − Sca-1 + c-Kit + HSPCs in coculture with OBN4 cells. ( a ) The increase in the number of non-adherent cells under coculture. LSK cells (10 5 cells per well of a 6-well plate) isolated as shown in Figure 4 were cocultured on monolayers of the OBN4 cells (6 × 10 4 cells per well) in the presence or absence of IL-7 (100 ngml −1 ) and SDF-1 (100 ngml −1 ) for 7 d. Black box, cytokine treatment. Open box, phosphate-buffered saline (PBS, non-cytokine treatment). ( b ) The increase in the number of B220 + cells under coculture. ( c ) In vitro B lymphopoiesis of HSPCs in coculture with OBN4 cells in the presence of IL-7 and SDF-1. The development of B-lineage cells was analyzed via flow cytometry with antibodies against B220, CD2, CD19, CD43, CD93, IgM and IgD during coculture with OBN4 for 7 d. The presented numbers indicate the percentage of each cell population. ( d ) The average numbers of B-lineage cells produced during coculture with OBN4 cells were summarized. NS, not significant. * P

    Article Snippet: Specific antibodies were purchased from the following commercial sources: anti-FLAG and anti-β-actin from Sigma-Aldrich; anti-B220, anti-Sca-1, anti-lineage cells (anti-Lin), anti-IgD and anti-IgM from BD Biosciences (San Jose, CA, USA); anti-Ki67, anti-Flk2, anti-CD2, anti-CD19, anti-CD23, anti-CD34, anti-CD43 and anti-CD93 from eBioscience (San Diego, CA, USA); and anti-receptor activator of nuclear factor κB ligand (RANKL) from Novus (Littleton, CO, USA).

    Techniques: In Vitro, Isolation, Flow Cytometry, Cytometry, Produced

    Osteoblastic characteristics of the OBN4 clone. ( a ) ALP activity of the OBN4 clone. ALP activities were analyzed between OBN4 and primary osteoblast (pOB) cells via stimulation with 50 μ M ascorbic acid, 10 m M β-glycerophosphate and 10 −7 M dexamethasone for 6 d. Photographs of ALP-positive cells in triplicate samples are presented (top panel), and ALP activities were quantified (bottom panel). ( b ) Mineralization of the OBN4 clone. The mineralization of cultured cells with 50 μ M ascorbic acid and 10 m M β-glycerophosphate for 21 d was measured using ARS staining. ARS-positive cells in triplicate samples were photographed (top panel), and ARS activities were quantified (bottom panel). ( c ) Expression of osteoblast markers in the OBN4 clone. The expression levels of osteocalcin (top panel) and osteopontin (bottom panel) were analyzed via real-time PCR analysis. ( d ) Expression of the Sca-1 lineage marker in the OBN4 clone. The expression of Sca-1 antigen was analyzed via flow cytometry with an anti-Sca-1 antibody (top panel). The percentages of the Sca-1 + cell population are summarized in the bottom panel. Mock, non-stained cells. ( e ) Comparison of the ability to support osteoclast differentiation in coculture assays. Cells were cocultured with bone marrow-derived HSCs in the presence of vitamin D3 and prostaglandin E2 for 6 d. Osteoclast formation was analyzed through TRAP staining (top panel). The number of TRAP-positive multinucleated osteoclasts (TRAP + MNCs (⩾three nuclei)) was counted (bottom panel). ( f ) RANKL expression in the OBN4 clone. The surface expression of RANKL was analyzed via flow cytometry with an anti-RANKL antibody (top panel). The percentages of the RANKL + cell population are summarized in the bottom panel. NS, not significant. * P

    Journal: Experimental & Molecular Medicine

    Article Title: Generation of an osteoblast-based artificial niche that supports in vitro B lymphopoiesis

    doi: 10.1038/emm.2017.189

    Figure Lengend Snippet: Osteoblastic characteristics of the OBN4 clone. ( a ) ALP activity of the OBN4 clone. ALP activities were analyzed between OBN4 and primary osteoblast (pOB) cells via stimulation with 50 μ M ascorbic acid, 10 m M β-glycerophosphate and 10 −7 M dexamethasone for 6 d. Photographs of ALP-positive cells in triplicate samples are presented (top panel), and ALP activities were quantified (bottom panel). ( b ) Mineralization of the OBN4 clone. The mineralization of cultured cells with 50 μ M ascorbic acid and 10 m M β-glycerophosphate for 21 d was measured using ARS staining. ARS-positive cells in triplicate samples were photographed (top panel), and ARS activities were quantified (bottom panel). ( c ) Expression of osteoblast markers in the OBN4 clone. The expression levels of osteocalcin (top panel) and osteopontin (bottom panel) were analyzed via real-time PCR analysis. ( d ) Expression of the Sca-1 lineage marker in the OBN4 clone. The expression of Sca-1 antigen was analyzed via flow cytometry with an anti-Sca-1 antibody (top panel). The percentages of the Sca-1 + cell population are summarized in the bottom panel. Mock, non-stained cells. ( e ) Comparison of the ability to support osteoclast differentiation in coculture assays. Cells were cocultured with bone marrow-derived HSCs in the presence of vitamin D3 and prostaglandin E2 for 6 d. Osteoclast formation was analyzed through TRAP staining (top panel). The number of TRAP-positive multinucleated osteoclasts (TRAP + MNCs (⩾three nuclei)) was counted (bottom panel). ( f ) RANKL expression in the OBN4 clone. The surface expression of RANKL was analyzed via flow cytometry with an anti-RANKL antibody (top panel). The percentages of the RANKL + cell population are summarized in the bottom panel. NS, not significant. * P

    Article Snippet: Specific antibodies were purchased from the following commercial sources: anti-FLAG and anti-β-actin from Sigma-Aldrich; anti-B220, anti-Sca-1, anti-lineage cells (anti-Lin), anti-IgD and anti-IgM from BD Biosciences (San Jose, CA, USA); anti-Ki67, anti-Flk2, anti-CD2, anti-CD19, anti-CD23, anti-CD34, anti-CD43 and anti-CD93 from eBioscience (San Diego, CA, USA); and anti-receptor activator of nuclear factor κB ligand (RANKL) from Novus (Littleton, CO, USA).

    Techniques: ALP Assay, Activity Assay, Cell Culture, Staining, Expressing, Real-time Polymerase Chain Reaction, Marker, Flow Cytometry, Cytometry, Derivative Assay

    CVB3 induced premature differentiation of cardiac progenitor cells. Three day-old mice were infected with eGFP-CVB3 (10 5 pfu IP) or mock-infected, and hearts were isolated at 2 days PI. Paraffin-embedded sections of heart tissue were deparaffinized and stained using antibodies against Sca-1, SM22, von Willebrand Factor (vWF), and sarcomeric actin (s-actin). (A) An increase in SM22 expression (red) was observed in infected heart sections, and Sca-1 + cells (green) expressed high levels of SM22 (light pink arrows). (B) vWF (red) was expressed near Sca-1 + capillaries following infection (light pink arrows). (C) In contrast, s-actin failed to co-localize with Sca-1 expression in the infected heart (light blue arrows). Representative images for three infected or mock-infected mice are shown. (D) c-kit + cells were isolated from the heart of juvenile mice, expanded in culture, and infected with eGFP-CVB3 (moi = 1). Relative expression levels of alpha actin and SM22 in mock-infected or CVB3-infected juvenile c-kit + cells were quantified by western analysis. A statistical significant difference was observed utilizing Student's T-test (**p

    Journal: PLoS Pathogens

    Article Title: The Impact of Juvenile Coxsackievirus Infection on Cardiac Progenitor Cells and Postnatal Heart Development

    doi: 10.1371/journal.ppat.1004249

    Figure Lengend Snippet: CVB3 induced premature differentiation of cardiac progenitor cells. Three day-old mice were infected with eGFP-CVB3 (10 5 pfu IP) or mock-infected, and hearts were isolated at 2 days PI. Paraffin-embedded sections of heart tissue were deparaffinized and stained using antibodies against Sca-1, SM22, von Willebrand Factor (vWF), and sarcomeric actin (s-actin). (A) An increase in SM22 expression (red) was observed in infected heart sections, and Sca-1 + cells (green) expressed high levels of SM22 (light pink arrows). (B) vWF (red) was expressed near Sca-1 + capillaries following infection (light pink arrows). (C) In contrast, s-actin failed to co-localize with Sca-1 expression in the infected heart (light blue arrows). Representative images for three infected or mock-infected mice are shown. (D) c-kit + cells were isolated from the heart of juvenile mice, expanded in culture, and infected with eGFP-CVB3 (moi = 1). Relative expression levels of alpha actin and SM22 in mock-infected or CVB3-infected juvenile c-kit + cells were quantified by western analysis. A statistical significant difference was observed utilizing Student's T-test (**p

    Article Snippet: For FACS analysis the following antibodies were used: PE-anti-Sca-1 (1∶100, BD #552108), FITC-anti-Sca-1 (1∶100, BD #557405), APC-anti-CD45 (1∶100, Caltag #MCD4505).

    Techniques: Mouse Assay, Infection, Isolation, Staining, Expressing, Western Blot

    CVB3 productively infected juvenile cardiac-derived c-kit+ cells grown in culture and in the juvenile heart. c-kit + cells were isolated from the heart of juvenile or adult mice and expanded in culture. (A) Cell cultures were infected with eGFP-CVB3 (moi = 1 or 1000), and infected cultures were followed by fluorescence microscopy for 10 days. (B) Viral titers were determined from supernatants of infected cell cultures over 10 days PI. (C) Juvenile c-kit + cells infected with eGFP-CVB3 (green) were fixed in 2% paraformaldehyde and stained by immunocytochemistry using an antibody against c-kit (red). (D) Juvenile and adult c-kit + cells were stained by immunocytochemistry using an antibody against coxsackie and adenovirus receptor (CAR, red). (E) Three day-old mice were infected with eGFP-CVB3 (10 5 pfu IP) or mock-infected, and hearts were isolated at 2 days PI. Paraffin-embedded sections of heart tissue were deparaffinized and stained using an antibody against c-kit (red) and GFP (virus, green). Many virus-infected c-kit + cells were observed in heart sections (white arrows). (F) Also, Sca-1 + cells (green) in heart tissue were shown to be infected with eGFP-CVB3 (red). Representative images of three infected mice are shown.

    Journal: PLoS Pathogens

    Article Title: The Impact of Juvenile Coxsackievirus Infection on Cardiac Progenitor Cells and Postnatal Heart Development

    doi: 10.1371/journal.ppat.1004249

    Figure Lengend Snippet: CVB3 productively infected juvenile cardiac-derived c-kit+ cells grown in culture and in the juvenile heart. c-kit + cells were isolated from the heart of juvenile or adult mice and expanded in culture. (A) Cell cultures were infected with eGFP-CVB3 (moi = 1 or 1000), and infected cultures were followed by fluorescence microscopy for 10 days. (B) Viral titers were determined from supernatants of infected cell cultures over 10 days PI. (C) Juvenile c-kit + cells infected with eGFP-CVB3 (green) were fixed in 2% paraformaldehyde and stained by immunocytochemistry using an antibody against c-kit (red). (D) Juvenile and adult c-kit + cells were stained by immunocytochemistry using an antibody against coxsackie and adenovirus receptor (CAR, red). (E) Three day-old mice were infected with eGFP-CVB3 (10 5 pfu IP) or mock-infected, and hearts were isolated at 2 days PI. Paraffin-embedded sections of heart tissue were deparaffinized and stained using an antibody against c-kit (red) and GFP (virus, green). Many virus-infected c-kit + cells were observed in heart sections (white arrows). (F) Also, Sca-1 + cells (green) in heart tissue were shown to be infected with eGFP-CVB3 (red). Representative images of three infected mice are shown.

    Article Snippet: For FACS analysis the following antibodies were used: PE-anti-Sca-1 (1∶100, BD #552108), FITC-anti-Sca-1 (1∶100, BD #557405), APC-anti-CD45 (1∶100, Caltag #MCD4505).

    Techniques: Infection, Derivative Assay, Isolation, Mouse Assay, Fluorescence, Microscopy, Staining, Immunocytochemistry

    Juvenile CVB3 infection triggered growth arrest in CPCs. Three day-old mice were infected with eGFP-CVB3 (10 5 pfu IP) or mock-infected, and hearts were isolated at 2 days PI. Paraffin-embedded sections of heart tissue were deparaffinized and stained using antibodies against Ki67, Sca-1, and c-kit. (A) Immunofluorescence microscopy revealed a sharp decline in Ki67 + cells in the neonatal myocardium at 2 days PI. (B) The decline of Ki67 + cells in the infected heart was quantified and shown to be statistically significant (*p

    Journal: PLoS Pathogens

    Article Title: The Impact of Juvenile Coxsackievirus Infection on Cardiac Progenitor Cells and Postnatal Heart Development

    doi: 10.1371/journal.ppat.1004249

    Figure Lengend Snippet: Juvenile CVB3 infection triggered growth arrest in CPCs. Three day-old mice were infected with eGFP-CVB3 (10 5 pfu IP) or mock-infected, and hearts were isolated at 2 days PI. Paraffin-embedded sections of heart tissue were deparaffinized and stained using antibodies against Ki67, Sca-1, and c-kit. (A) Immunofluorescence microscopy revealed a sharp decline in Ki67 + cells in the neonatal myocardium at 2 days PI. (B) The decline of Ki67 + cells in the infected heart was quantified and shown to be statistically significant (*p

    Article Snippet: For FACS analysis the following antibodies were used: PE-anti-Sca-1 (1∶100, BD #552108), FITC-anti-Sca-1 (1∶100, BD #557405), APC-anti-CD45 (1∶100, Caltag #MCD4505).

    Techniques: Infection, Mouse Assay, Isolation, Staining, Immunofluorescence, Microscopy

    Acute CVB3 infection in juvenile mice induced an increase in myocardial Sca-1 expression. Three day-old mice were infected with eGFP-CVB3 (10 5 pfu IP) or mock-infected, and hearts were isolated at 2 days PI. Paraffin-embedded sections of heart tissue were deparaffinized and stained using antibodies against Sca-1. Alternatively, hearts were mechanically dissociated and cells were fixed in 10% formalin in PBS for FACS analysis (A) An increase in Sca-1 + cells (green) was observed in the heart following infection. (B) FACS analysis confirmed an elevation of Sca-1 + cells in the heart following infection. Primarily resident Sca-1 + , but also blood-derived (CD45 + ) Sca-1 + cells were shown to increase in number in the heart following infection. Representative flow cytometric results for three infected or mock-infected mice are shown.

    Journal: PLoS Pathogens

    Article Title: The Impact of Juvenile Coxsackievirus Infection on Cardiac Progenitor Cells and Postnatal Heart Development

    doi: 10.1371/journal.ppat.1004249

    Figure Lengend Snippet: Acute CVB3 infection in juvenile mice induced an increase in myocardial Sca-1 expression. Three day-old mice were infected with eGFP-CVB3 (10 5 pfu IP) or mock-infected, and hearts were isolated at 2 days PI. Paraffin-embedded sections of heart tissue were deparaffinized and stained using antibodies against Sca-1. Alternatively, hearts were mechanically dissociated and cells were fixed in 10% formalin in PBS for FACS analysis (A) An increase in Sca-1 + cells (green) was observed in the heart following infection. (B) FACS analysis confirmed an elevation of Sca-1 + cells in the heart following infection. Primarily resident Sca-1 + , but also blood-derived (CD45 + ) Sca-1 + cells were shown to increase in number in the heart following infection. Representative flow cytometric results for three infected or mock-infected mice are shown.

    Article Snippet: For FACS analysis the following antibodies were used: PE-anti-Sca-1 (1∶100, BD #552108), FITC-anti-Sca-1 (1∶100, BD #557405), APC-anti-CD45 (1∶100, Caltag #MCD4505).

    Techniques: Infection, Mouse Assay, Expressing, Isolation, Staining, FACS, Derivative Assay, Flow Cytometry

    TS cell markers, Cdx2 and Esrrb were more highly expressed in Sca-1 + trophoblast isolated from differentiated TS cell cultures than in non-sorted, proliferating TS cell cultures ( A ). Sca-1 + trophoblast isolated from differentiated TS cultures gave rise to proliferating trophoblast cell lines that when differentiated by the removal of growth factors, expressed genes indicative of syncytiotrophoblast ( Gcm1, Syna ), spongiotrophoblast ( Tpbpa ) and TGCs ( Prl3b1 ) ( B ). These differentiated cells were also positive for alkaline phosphatase activity ( C , blue) and PAS ( D , magenta/arrowhead) indicating differentiation of sinusoidal TGCs and glycogen trophoblast, respectively. D0 represents undifferentiated, proliferating TS cells while, D2, D4, D6 represent cultures differentiated for 2, 4 or 6 days. In C, scale bar = 500 um. In D, scale bar = 100 um.

    Journal: Scientific Reports

    Article Title: Sca-1 identifies a trophoblast population with multipotent potential in the mid-gestation mouse placenta

    doi: 10.1038/s41598-017-06008-2

    Figure Lengend Snippet: TS cell markers, Cdx2 and Esrrb were more highly expressed in Sca-1 + trophoblast isolated from differentiated TS cell cultures than in non-sorted, proliferating TS cell cultures ( A ). Sca-1 + trophoblast isolated from differentiated TS cultures gave rise to proliferating trophoblast cell lines that when differentiated by the removal of growth factors, expressed genes indicative of syncytiotrophoblast ( Gcm1, Syna ), spongiotrophoblast ( Tpbpa ) and TGCs ( Prl3b1 ) ( B ). These differentiated cells were also positive for alkaline phosphatase activity ( C , blue) and PAS ( D , magenta/arrowhead) indicating differentiation of sinusoidal TGCs and glycogen trophoblast, respectively. D0 represents undifferentiated, proliferating TS cells while, D2, D4, D6 represent cultures differentiated for 2, 4 or 6 days. In C, scale bar = 500 um. In D, scale bar = 100 um.

    Article Snippet: Anti-Sca-1 antibodies were from BD Biosciences (557403, clone D7) and Miltenyi (130-102-297, clone D7).

    Techniques: Isolation, Activity Assay

    Upon removal of FGF4, mouse TS cell cultures were initially proliferative but increased in number only until day 4 of differentiation ( A ). Sca-1 + trophoblast cells isolated from non-proliferating, differentiated TS cultures and plated at decreasing densities in conditions to support TS proliferation formed colonies however Sca-1 − and non-sorted, differentiated TS cells did not ( B ). When these cells were compared to the Sca-1 − subpopulation of trophoblast cells by qRT-PCR, Sca-1 + positive cells displayed higher expression of Sca-1 and the trophoblast stem and progenitor markers, Esrrb and Epcam, Rhox4 , respectively, while Ascl2 was not different ( C ).

    Journal: Scientific Reports

    Article Title: Sca-1 identifies a trophoblast population with multipotent potential in the mid-gestation mouse placenta

    doi: 10.1038/s41598-017-06008-2

    Figure Lengend Snippet: Upon removal of FGF4, mouse TS cell cultures were initially proliferative but increased in number only until day 4 of differentiation ( A ). Sca-1 + trophoblast cells isolated from non-proliferating, differentiated TS cultures and plated at decreasing densities in conditions to support TS proliferation formed colonies however Sca-1 − and non-sorted, differentiated TS cells did not ( B ). When these cells were compared to the Sca-1 − subpopulation of trophoblast cells by qRT-PCR, Sca-1 + positive cells displayed higher expression of Sca-1 and the trophoblast stem and progenitor markers, Esrrb and Epcam, Rhox4 , respectively, while Ascl2 was not different ( C ).

    Article Snippet: Anti-Sca-1 antibodies were from BD Biosciences (557403, clone D7) and Miltenyi (130-102-297, clone D7).

    Techniques: Isolation, Quantitative RT-PCR, Expressing

    Cartoon depicting proposed position of Sca-1 + cells within the mouse trophoblast stem cell lineage. P-TGC, parietal trophoblast giant cell; SpA-TGC, spiral artery-associated trophoblast giant cell; Gly-T, glycogen trophoblast; Sp-T, spongiotrophoblast; S-TGC, sinusoidal trophoblast giant cell; Syn-T, syncytiotrophoblast.

    Journal: Scientific Reports

    Article Title: Sca-1 identifies a trophoblast population with multipotent potential in the mid-gestation mouse placenta

    doi: 10.1038/s41598-017-06008-2

    Figure Lengend Snippet: Cartoon depicting proposed position of Sca-1 + cells within the mouse trophoblast stem cell lineage. P-TGC, parietal trophoblast giant cell; SpA-TGC, spiral artery-associated trophoblast giant cell; Gly-T, glycogen trophoblast; Sp-T, spongiotrophoblast; S-TGC, sinusoidal trophoblast giant cell; Syn-T, syncytiotrophoblast.

    Article Snippet: Anti-Sca-1 antibodies were from BD Biosciences (557403, clone D7) and Miltenyi (130-102-297, clone D7).

    Techniques:

    Sca-1 + trophoblast isolated from differentiated TS cell cultures were re-plated in TS media plus growth factors and cultured in 5% CO 2 in air (Normoxia) or 1% O 2 (Hypoxia) for 48 hours. These cultures were compared to proliferating, non-sorted TS cells plated in the same conditions and at the same seeding density (TS). Hypoxia promoted colony formation and proliferation compared to normoxia ( A ). After 48 hours of culture in hypoxia in the presence of growth factors, expression of Cdx2 and Esrrb in Sca-1 + trophoblast was maintained, while Eomes expression was up regulated in these cells as detected by qRT-PCR ( B ). Cells were stained with hematoxylin and eosin. Scale bar = 500 um.

    Journal: Scientific Reports

    Article Title: Sca-1 identifies a trophoblast population with multipotent potential in the mid-gestation mouse placenta

    doi: 10.1038/s41598-017-06008-2

    Figure Lengend Snippet: Sca-1 + trophoblast isolated from differentiated TS cell cultures were re-plated in TS media plus growth factors and cultured in 5% CO 2 in air (Normoxia) or 1% O 2 (Hypoxia) for 48 hours. These cultures were compared to proliferating, non-sorted TS cells plated in the same conditions and at the same seeding density (TS). Hypoxia promoted colony formation and proliferation compared to normoxia ( A ). After 48 hours of culture in hypoxia in the presence of growth factors, expression of Cdx2 and Esrrb in Sca-1 + trophoblast was maintained, while Eomes expression was up regulated in these cells as detected by qRT-PCR ( B ). Cells were stained with hematoxylin and eosin. Scale bar = 500 um.

    Article Snippet: Anti-Sca-1 antibodies were from BD Biosciences (557403, clone D7) and Miltenyi (130-102-297, clone D7).

    Techniques: Isolation, Cell Culture, Expressing, Quantitative RT-PCR, Staining

    Sca-1 + and Epcam + cells isolated from mouse placentae at E11.5 expressed the trophoblast marker, Krt8 by qRT-PCR. These cells also expressed other known markers of trophoblast, including Cdx2, Eomes and Epcam . Epcam was expressed in both Sca-1 + and Epcam + cells however Sca-1 was only detected in Sca-1 + cells. The syncytiotrophoblast marker, Gcm1 , was detected only in Epcam + cells.

    Journal: Scientific Reports

    Article Title: Sca-1 identifies a trophoblast population with multipotent potential in the mid-gestation mouse placenta

    doi: 10.1038/s41598-017-06008-2

    Figure Lengend Snippet: Sca-1 + and Epcam + cells isolated from mouse placentae at E11.5 expressed the trophoblast marker, Krt8 by qRT-PCR. These cells also expressed other known markers of trophoblast, including Cdx2, Eomes and Epcam . Epcam was expressed in both Sca-1 + and Epcam + cells however Sca-1 was only detected in Sca-1 + cells. The syncytiotrophoblast marker, Gcm1 , was detected only in Epcam + cells.

    Article Snippet: Anti-Sca-1 antibodies were from BD Biosciences (557403, clone D7) and Miltenyi (130-102-297, clone D7).

    Techniques: Isolation, Marker, Quantitative RT-PCR

    Sca-1 + cells isolated from E11.5 placentae formed proliferating, epithelial-like colonies that could be repeatedly passaged ( A ). When analyzed by RT-PCR, three independent Sca-1-derived colonies expressed genes indicative of trophoblast stem cells ( Cdx2, Esrrb ), syncytiotrophoblast ( Gcm1 ) and trophoblast giant cells ( Prl3d1, Prl3b1 ) ( B ). The frequency of pHH3 + /cytokeratin + trophoblast was increased in the labyrinth layer in response to RUPP ( C ). The expression patterns of Eomes ( D ) and Sca-1 ( F ) were expanded in RUPP placentae ( D,F ) when compared to sham-operated controls ( E , Eomes ; G , Sca-1 ). U and D reflect undifferentiated and differentiated cultures respectively. Scale bar = 100 um. RT-PCR results presented in ( B ) are cropped and organized to be shown as a composite from multiple gel pictures.

    Journal: Scientific Reports

    Article Title: Sca-1 identifies a trophoblast population with multipotent potential in the mid-gestation mouse placenta

    doi: 10.1038/s41598-017-06008-2

    Figure Lengend Snippet: Sca-1 + cells isolated from E11.5 placentae formed proliferating, epithelial-like colonies that could be repeatedly passaged ( A ). When analyzed by RT-PCR, three independent Sca-1-derived colonies expressed genes indicative of trophoblast stem cells ( Cdx2, Esrrb ), syncytiotrophoblast ( Gcm1 ) and trophoblast giant cells ( Prl3d1, Prl3b1 ) ( B ). The frequency of pHH3 + /cytokeratin + trophoblast was increased in the labyrinth layer in response to RUPP ( C ). The expression patterns of Eomes ( D ) and Sca-1 ( F ) were expanded in RUPP placentae ( D,F ) when compared to sham-operated controls ( E , Eomes ; G , Sca-1 ). U and D reflect undifferentiated and differentiated cultures respectively. Scale bar = 100 um. RT-PCR results presented in ( B ) are cropped and organized to be shown as a composite from multiple gel pictures.

    Article Snippet: Anti-Sca-1 antibodies were from BD Biosciences (557403, clone D7) and Miltenyi (130-102-297, clone D7).

    Techniques: Isolation, Reverse Transcription Polymerase Chain Reaction, Derivative Assay, Expressing

    Sca-1 + trophoblast isolated from differentiated TS cell cultures and re-plated in TS media plus growth factors formed colonies comparable to TS cells plated at the same density from proliferating cultures (compare TS and Sca-1 + , Day 2). After six days of culture in TS media plus growth factors, both Sca-1 + and TS cells were confluent and appeared similar in morphology (compare TS and Sca-1 + , Day 6). Cells were stained with hematoxylin and eosin. Scale bar = 500 um.

    Journal: Scientific Reports

    Article Title: Sca-1 identifies a trophoblast population with multipotent potential in the mid-gestation mouse placenta

    doi: 10.1038/s41598-017-06008-2

    Figure Lengend Snippet: Sca-1 + trophoblast isolated from differentiated TS cell cultures and re-plated in TS media plus growth factors formed colonies comparable to TS cells plated at the same density from proliferating cultures (compare TS and Sca-1 + , Day 2). After six days of culture in TS media plus growth factors, both Sca-1 + and TS cells were confluent and appeared similar in morphology (compare TS and Sca-1 + , Day 6). Cells were stained with hematoxylin and eosin. Scale bar = 500 um.

    Article Snippet: Anti-Sca-1 antibodies were from BD Biosciences (557403, clone D7) and Miltenyi (130-102-297, clone D7).

    Techniques: Isolation, Staining

    Epcam + trophoblast isolated from TS cell cultures and re-plated in TS media containing growth factors to promote proliferation formed small colonies after 2 days of culture but did not proliferate ( A , compare Day 2 to Day 6). After 6 days in culture in TS media plus growth factors, the cells had not expanded and had the morphology of differentiated trophoblast, with large nuclei and cell bodies (compare Day 6 to Fig. 3 , Day 2). Sca-1 + cells compared to Epcam + cells displayed increased expression of the TS cell marker, Esrrb while Epcam + cells expressed the syncytiotrophoblast marker, Gcm1 but not Sca-1 or Esrrb ( B ). Insets in ( A ) show morphology of plated Epcam + trophoblast cells at high magnification (400x). Cells were stained with hematoxylin and eosin. Scale bar = 500 um.

    Journal: Scientific Reports

    Article Title: Sca-1 identifies a trophoblast population with multipotent potential in the mid-gestation mouse placenta

    doi: 10.1038/s41598-017-06008-2

    Figure Lengend Snippet: Epcam + trophoblast isolated from TS cell cultures and re-plated in TS media containing growth factors to promote proliferation formed small colonies after 2 days of culture but did not proliferate ( A , compare Day 2 to Day 6). After 6 days in culture in TS media plus growth factors, the cells had not expanded and had the morphology of differentiated trophoblast, with large nuclei and cell bodies (compare Day 6 to Fig. 3 , Day 2). Sca-1 + cells compared to Epcam + cells displayed increased expression of the TS cell marker, Esrrb while Epcam + cells expressed the syncytiotrophoblast marker, Gcm1 but not Sca-1 or Esrrb ( B ). Insets in ( A ) show morphology of plated Epcam + trophoblast cells at high magnification (400x). Cells were stained with hematoxylin and eosin. Scale bar = 500 um.

    Article Snippet: Anti-Sca-1 antibodies were from BD Biosciences (557403, clone D7) and Miltenyi (130-102-297, clone D7).

    Techniques: Isolation, Expressing, Marker, Staining

    Sca-1 was determined to be highly expressed in undifferentiated TS cells in culture by Northern blot and qRT-PCR ( A ) and FACS analysis ( B ). Day 0/0 represents undifferentiated, proliferating TS cells while 2, 4, 6 and Day 2, Day 4 and Day 6 represent cultures differentiated for 2, 4 or 6 days. Sca-1 was also expressed in TS cells in culture as detected by immunofluorescence ( C ; +FGF) however became undetectable following differentiation ( C ; -FGF). Immunofluorescence in cells ( C ) in the presence and absence of FGF matched the pattern of expression observed by FACS ( B ), however was not obvious in as a high frequency (+FGF) which is likely due to the more sensitive detection of Sca-1 expression by FACS. Scale bar = 100 um. Northern blot results shown in ( A ) are cropped and organized to be presented as a composite from multiple blots/films.

    Journal: Scientific Reports

    Article Title: Sca-1 identifies a trophoblast population with multipotent potential in the mid-gestation mouse placenta

    doi: 10.1038/s41598-017-06008-2

    Figure Lengend Snippet: Sca-1 was determined to be highly expressed in undifferentiated TS cells in culture by Northern blot and qRT-PCR ( A ) and FACS analysis ( B ). Day 0/0 represents undifferentiated, proliferating TS cells while 2, 4, 6 and Day 2, Day 4 and Day 6 represent cultures differentiated for 2, 4 or 6 days. Sca-1 was also expressed in TS cells in culture as detected by immunofluorescence ( C ; +FGF) however became undetectable following differentiation ( C ; -FGF). Immunofluorescence in cells ( C ) in the presence and absence of FGF matched the pattern of expression observed by FACS ( B ), however was not obvious in as a high frequency (+FGF) which is likely due to the more sensitive detection of Sca-1 expression by FACS. Scale bar = 100 um. Northern blot results shown in ( A ) are cropped and organized to be presented as a composite from multiple blots/films.

    Article Snippet: Anti-Sca-1 antibodies were from BD Biosciences (557403, clone D7) and Miltenyi (130-102-297, clone D7).

    Techniques: Northern Blot, Quantitative RT-PCR, FACS, Immunofluorescence, Expressing

    Sca-1 was expressed in the chorion of the mouse placenta at E8.5 ( A ; arrow). The inset in ( A ) is shown at higher magnification in ( B ) and from a serial section in ( C ), where protein expression is indicated by arrows. Controls in which no primary antibody was used showed no positive staining (not shown). Sca-1 was expressed in the labyrinth at E10.5 ( D ) and E16.5 ( E ). The inset in ( E ) shows Sca-1 expression (arrowhead, blue; ISH) in cytokeratin positive trophoblast of the labyrinth (brown; IHC). Epcam mRNA expression was also detected in the chorion and appeared to overlap with Sca-1 expression at E8.5 ( F ). At E10.5, Epcam expression was more broadly expressed in the labyrinth than Sca-1 ( G , compare with D ) but by E16.5, both Epcam mRNA and protein were appreciably reduced however still present in more cells in the labyrinth than Sca-1 ( H , I , compare with E ). ch, chorion; mes, embryonic mesoderm; lab, labyrinth. Scale bar = 100 um.

    Journal: Scientific Reports

    Article Title: Sca-1 identifies a trophoblast population with multipotent potential in the mid-gestation mouse placenta

    doi: 10.1038/s41598-017-06008-2

    Figure Lengend Snippet: Sca-1 was expressed in the chorion of the mouse placenta at E8.5 ( A ; arrow). The inset in ( A ) is shown at higher magnification in ( B ) and from a serial section in ( C ), where protein expression is indicated by arrows. Controls in which no primary antibody was used showed no positive staining (not shown). Sca-1 was expressed in the labyrinth at E10.5 ( D ) and E16.5 ( E ). The inset in ( E ) shows Sca-1 expression (arrowhead, blue; ISH) in cytokeratin positive trophoblast of the labyrinth (brown; IHC). Epcam mRNA expression was also detected in the chorion and appeared to overlap with Sca-1 expression at E8.5 ( F ). At E10.5, Epcam expression was more broadly expressed in the labyrinth than Sca-1 ( G , compare with D ) but by E16.5, both Epcam mRNA and protein were appreciably reduced however still present in more cells in the labyrinth than Sca-1 ( H , I , compare with E ). ch, chorion; mes, embryonic mesoderm; lab, labyrinth. Scale bar = 100 um.

    Article Snippet: Anti-Sca-1 antibodies were from BD Biosciences (557403, clone D7) and Miltenyi (130-102-297, clone D7).

    Techniques: Expressing, Staining, In Situ Hybridization, Immunohistochemistry

    Flow cytometric analyses of CPC cells for expression of the cell surface markers Sca-1, CD31, c-kit, Flk-1, CD105 and CD45.

    Journal: PLoS ONE

    Article Title: Infrared Fluorescent Protein 1.4 Genetic Labeling Tracks Engrafted Cardiac Progenitor Cells in Mouse Ischemic Hearts

    doi: 10.1371/journal.pone.0107841

    Figure Lengend Snippet: Flow cytometric analyses of CPC cells for expression of the cell surface markers Sca-1, CD31, c-kit, Flk-1, CD105 and CD45.

    Article Snippet: Briefly, CPC cells were blocked with 5% rat serum and stained with a panel of conjugated antibodies, including anti-Sca-1-PE, anti-CD31-biotin,, anti-c-kit-biotin, anti-CD45-PE-cy7, anti-Flk1-PerCP-cy5.5 and anti-CD105-V450 (BD Biosciences, San Jose, CA), or isotype-matched control antibodies.

    Techniques: Flow Cytometry, Expressing

    The change in the spatial distribution of H4K16ac upon aging is linked to a change in HSC function. a ]. b Representative FACS dot plots of HSCs (Lin-c-kit+Sca-1+Flk2-CD34-), ST-HSCs (Lin-c-kit+Sca-1+Flk2-CD34+), LMPPs (Lin-c-kit+Sca-1+Flk2+CD34+), and LSKs (Lin-c-kit+Sca-1+) gating strategy of young and aged lineage-depleted BM cells. c Percentage of young and aged HSCs with a polar distribution of H4K16ac, H4K8ac, H4K5ac, H3K27ac, H3K4me1, and H3K4me3; n = 3–4 biological repeats; ~ 150–200 single HSCs scored per sample in total; * p

    Journal: Genome Biology

    Article Title: LaminA/C regulates epigenetic and chromatin architecture changes upon aging of hematopoietic stem cells

    doi: 10.1186/s13059-018-1557-3

    Figure Lengend Snippet: The change in the spatial distribution of H4K16ac upon aging is linked to a change in HSC function. a ]. b Representative FACS dot plots of HSCs (Lin-c-kit+Sca-1+Flk2-CD34-), ST-HSCs (Lin-c-kit+Sca-1+Flk2-CD34+), LMPPs (Lin-c-kit+Sca-1+Flk2+CD34+), and LSKs (Lin-c-kit+Sca-1+) gating strategy of young and aged lineage-depleted BM cells. c Percentage of young and aged HSCs with a polar distribution of H4K16ac, H4K8ac, H4K5ac, H3K27ac, H3K4me1, and H3K4me3; n = 3–4 biological repeats; ~ 150–200 single HSCs scored per sample in total; * p

    Article Snippet: After lineage depletion by magnetic separation (Dynalbeads, Invitrogen), cells were stained with anti-Sca-1 (clone D7) (eBioscience), anti-c-kit (clone 2B8) (eBioscience), anti-CD34 (clone RAM34) (eBioscience), anti-CD127 (clone A7R34) (eBioscience), anti-Flk-2 (clone A2F10) (eBioscience), and streptavidin (eBioscience).

    Techniques: FACS

    Conditional deletion of CTC1 . ( A ) Schematic showing CTC1 genomic locus, targeting construct and CTC1 null allele. Black boxes, coding exons, white box, non-coding exon; red box, exon 6; arrowheads, loxP sites; green rectangle, PGK-neo gene; EV, EcoRV; blue rectangles, left and right arm probes for genomic Southern. ( B ) Kaplan–Meier survival curve of WT and CTC1 null mice. Number of mice analysed is indicated. ( C ) Weight of spleens from WT and CTC1 null mice. P =0.0013. Two-tailed t -test was used to calculate statistical significance. ( D ) Histology of femurs derived from 25-day-old WT and CTC1 null mice. Scale bar, 100 μm. ( E ) Top: Expression of Sca-1 and c-Kit in multilineage negative BM cells from WT and CTC1 −/− mice. Bottom: BrdU/7-AAD FACS profiles of BM LSK cells (Lin − Sca-1 + c-kit + ) derived from aged WT and CTC1 −/− mice. Results were expressed as percentage of total nucleated BM cells for each fraction. Number of mice analysed is indicated.

    Journal: The EMBO Journal

    Article Title: CTC1 deletion results in defective telomere replication, leading to catastrophic telomere loss and stem cell exhaustion

    doi: 10.1038/emboj.2012.96

    Figure Lengend Snippet: Conditional deletion of CTC1 . ( A ) Schematic showing CTC1 genomic locus, targeting construct and CTC1 null allele. Black boxes, coding exons, white box, non-coding exon; red box, exon 6; arrowheads, loxP sites; green rectangle, PGK-neo gene; EV, EcoRV; blue rectangles, left and right arm probes for genomic Southern. ( B ) Kaplan–Meier survival curve of WT and CTC1 null mice. Number of mice analysed is indicated. ( C ) Weight of spleens from WT and CTC1 null mice. P =0.0013. Two-tailed t -test was used to calculate statistical significance. ( D ) Histology of femurs derived from 25-day-old WT and CTC1 null mice. Scale bar, 100 μm. ( E ) Top: Expression of Sca-1 and c-Kit in multilineage negative BM cells from WT and CTC1 −/− mice. Bottom: BrdU/7-AAD FACS profiles of BM LSK cells (Lin − Sca-1 + c-kit + ) derived from aged WT and CTC1 −/− mice. Results were expressed as percentage of total nucleated BM cells for each fraction. Number of mice analysed is indicated.

    Article Snippet: In addition, the cocktail also contained anti-Sca-1-PE (12-5981 eBioscience) and anti-c-Kit-APC (17-1171 eBioscience).

    Techniques: Construct, Mouse Assay, Two Tailed Test, Derivative Assay, Expressing, FACS

    Y-27632 treatment increases the prostate colony-forming activity of LSC (Lin − Sca-1 + CD49f high ) basal cells by 8 fold. FACS sorted LSC cells from dsRED mice were plated on mitomysin-treated Swiss3T3 layer cells in a dilution series ranging from 133 to 2,667 cells per well with and without Y-27632. Graph shows the number of colonies grown at each dilution plotted versus the input cell number. The slopes represent colony-forming units.

    Journal: PLoS ONE

    Article Title: ROCK Inhibitor Y-27632 Suppresses Dissociation-Induced Apoptosis of Murine Prostate Stem/Progenitor Cells and Increases Their Cloning Efficiency

    doi: 10.1371/journal.pone.0018271

    Figure Lengend Snippet: Y-27632 treatment increases the prostate colony-forming activity of LSC (Lin − Sca-1 + CD49f high ) basal cells by 8 fold. FACS sorted LSC cells from dsRED mice were plated on mitomysin-treated Swiss3T3 layer cells in a dilution series ranging from 133 to 2,667 cells per well with and without Y-27632. Graph shows the number of colonies grown at each dilution plotted versus the input cell number. The slopes represent colony-forming units.

    Article Snippet: The antibodies used were biotin- or FITC-anti CD31,CD45 and Ter119 antibodies (eBioscience, San Diego, CA), FITC- or PE-anti Sca-1 antibody (eBioscience, San Diego, CA), Alexa 647-anti CD49f antibody (Biolegend, San Diego, CA) and strepavidin-Alexa 750 (Invitrogen, Carlsbad, CA).

    Techniques: Activity Assay, FACS, Mouse Assay

    Characterization of MSC mix and NCSC mix isolated from the bone marrow of adult Wnt1-CRE/R26R-LacZ mice. After recombination, NCSCs from Wnt1-CRE/R26R-LacZ mice express LacZ gene. MSCs did not undergo Cre/Lox recombination and conserved the PGK-Neo cassette ( a ). MSC mix are adherent fibroblast-like cells, do not express β-galactosidase ( b ) or Sox2 c (red), slightly express Nestin ( c ) (green), p75NTR ( d ) (red), and Sca-1 ( d ) (green). NCSC mix have a similar morphology, express β-galactosidase ( e ), Nestin ( f ) (green), Sox2 ( f ) (red) and p75NTR ( g ) red), but not Sca-1( g ) (green). Scale bar = 20 μm. MSC mesenchymal stem cell, NCSC neural crest stem cell

    Journal: Stem Cell Research & Therapy

    Article Title: Adult bone marrow mesenchymal and neural crest stem cells are chemoattractive and accelerate motor recovery in a mouse model of spinal cord injury

    doi: 10.1186/s13287-015-0202-2

    Figure Lengend Snippet: Characterization of MSC mix and NCSC mix isolated from the bone marrow of adult Wnt1-CRE/R26R-LacZ mice. After recombination, NCSCs from Wnt1-CRE/R26R-LacZ mice express LacZ gene. MSCs did not undergo Cre/Lox recombination and conserved the PGK-Neo cassette ( a ). MSC mix are adherent fibroblast-like cells, do not express β-galactosidase ( b ) or Sox2 c (red), slightly express Nestin ( c ) (green), p75NTR ( d ) (red), and Sca-1 ( d ) (green). NCSC mix have a similar morphology, express β-galactosidase ( e ), Nestin ( f ) (green), Sox2 ( f ) (red) and p75NTR ( g ) red), but not Sca-1( g ) (green). Scale bar = 20 μm. MSC mesenchymal stem cell, NCSC neural crest stem cell

    Article Snippet: For specific immunofluorescent staining, anti-nestin (1:300, NB100-1604, Novus Biologicals, Littleton, CO, USA), anti-Sox2 (1:250; sc-17320, Santa Cruz, Dallas, Texas, USA), anti p75NTR (1:200, AB1554, Millipore, Darmstadt, Germany), anti-Sca-1 (1:100, ab25195, Abcam, Cambridge, United Kingdom), anti-arginase 1 (1:100, 610708 BD Biosciences, Franklin Lakes, NJ, USA), anti-Iba1 (1:800, 019-19741, Wako Chemicals, Richmond, VA, USA), anti-GFAP (1:1000, 20334, Dako, Glostrup, Denmark), anti-laminin (1:200, L9393, Sigma-Aldrich, Saint-Louis, MO, USA) were diluted in PBS 0.1 M overnight at 4 °C.

    Techniques: Isolation, Mouse Assay

    hCMPC in the human heart. (A-G) Immunohistochemistry for Sca-1 in foetal and adult heart. (B, C) High power magnification of areas in A. (D) Atrial ventricular boundary. (E) High power magnification of area in D. (F, H) hCMPCs in biopsy from adult patient. (G) High power magnification of area in F. (I) IgG control. Arrows designate some of the hCMPCs. Magnification: A, D, F, H, I: 100x, B, C, E, G: 200x.

    Journal: Netherlands Heart Journal

    Article Title: Progenitor cells isolated from the human heart: a potential cell source for regenerative therapy

    doi:

    Figure Lengend Snippet: hCMPC in the human heart. (A-G) Immunohistochemistry for Sca-1 in foetal and adult heart. (B, C) High power magnification of areas in A. (D) Atrial ventricular boundary. (E) High power magnification of area in D. (F, H) hCMPCs in biopsy from adult patient. (G) High power magnification of area in F. (I) IgG control. Arrows designate some of the hCMPCs. Magnification: A, D, F, H, I: 100x, B, C, E, G: 200x.

    Article Snippet: The sections were incubated with the anti-Sca-1 antibody (Pharmingen), diluted 1:100 in blocking solution, o/n at 4°C.

    Techniques: Immunohistochemistry

    (a) Culture of muscle-derived stem cells. MDSCs were cultured and subcultured at confluent of 80%. Bright view images were taken at day 1 and day 5 after subconfluent. (b) Immunohistochemical staining of Sca-1. MDSCs were subcultured. Expression of Sca-1 was assessed by immunohistochemical staining analysis at day 2 using an anti-Sca-1 antibody (Wuhan Boster Biological Technology Co., Ltd.).

    Journal: Journal of Biomedicine and Biotechnology

    Article Title: BMP9-Induced Osteogenetic Differentiation and Bone Formation of Muscle-Derived Stem Cells

    doi: 10.1155/2012/610952

    Figure Lengend Snippet: (a) Culture of muscle-derived stem cells. MDSCs were cultured and subcultured at confluent of 80%. Bright view images were taken at day 1 and day 5 after subconfluent. (b) Immunohistochemical staining of Sca-1. MDSCs were subcultured. Expression of Sca-1 was assessed by immunohistochemical staining analysis at day 2 using an anti-Sca-1 antibody (Wuhan Boster Biological Technology Co., Ltd.).

    Article Snippet: Anti-Sca-1 antibody (Wuhan Boster Biological Technology Co., Ltd.) and nanohydroxyapatite/polyamide bone cement were provided by Research Center of Nano-biomaterials, Sichuan University.

    Techniques: Derivative Assay, Cell Culture, Immunohistochemistry, Staining, Expressing

    Prolonged expression of Sox17 results in formation of bronchiolar-like structures in the alveoli. Adult CCSPrtTA/tetO-Sox17 transgenic mice were maintained on Dox for 12 months. (A) H E staining shows the presence of bronchiolar-like sheets of cells in the peripheral lung. Arrowhead indicates the pleural surface. (B–D) Immunostaining for Sox17 (B), Foxj1 (C), and CCSP (D) was performed on lung sections. The Sox17-induced bronchiolar-like structures contained cells expressing proximal airway markers CCSP and Foxj1. (E–H) Immunofluorescent staining for CCSP (E), proSP-C (F), and Sca-1 (G). The bronchiolar-like structures (dotted outline) contained cells that express Sca-1 and subsets of cells expressing CCSP or proSP-C. Arrowheads demark CCSP-expressing cells in the bronchiolar epithelium and the arrow indicates normal proSP-C expression in a type II cell. Nuclei are stained with DAPI (H; blue) Scale bars, 50 µm.

    Journal: PLoS ONE

    Article Title: Sox17 Promotes Cell Cycle Progression and Inhibits TGF-?/Smad3 Signaling to Initiate Progenitor Cell Behavior in the Respiratory Epithelium

    doi: 10.1371/journal.pone.0005711

    Figure Lengend Snippet: Prolonged expression of Sox17 results in formation of bronchiolar-like structures in the alveoli. Adult CCSPrtTA/tetO-Sox17 transgenic mice were maintained on Dox for 12 months. (A) H E staining shows the presence of bronchiolar-like sheets of cells in the peripheral lung. Arrowhead indicates the pleural surface. (B–D) Immunostaining for Sox17 (B), Foxj1 (C), and CCSP (D) was performed on lung sections. The Sox17-induced bronchiolar-like structures contained cells expressing proximal airway markers CCSP and Foxj1. (E–H) Immunofluorescent staining for CCSP (E), proSP-C (F), and Sca-1 (G). The bronchiolar-like structures (dotted outline) contained cells that express Sca-1 and subsets of cells expressing CCSP or proSP-C. Arrowheads demark CCSP-expressing cells in the bronchiolar epithelium and the arrow indicates normal proSP-C expression in a type II cell. Nuclei are stained with DAPI (H; blue) Scale bars, 50 µm.

    Article Snippet: Immunohistochemistry was performed using primary antibodies for guinea pig-anti Sox17 (1∶10,000), rabbit anti-phospho-histone H3 (1∶1000; Santa Cruz), rabbit anti-CCSP (1∶5000), rabbit anti-Foxj1 (1∶14,000), rabbit anti-cyclin D1 (1∶400; Abcam), and rat anti-Sca-1 (1∶250; BD Pharmingen).

    Techniques: Expressing, Transgenic Assay, Mouse Assay, Staining, Immunostaining

    Sox17 increases Sca-1 expression adult respiratory epithelial cells. Adult CCSPrtTA control (A) and CCSPrtTA/tetO-Sox17 (B–N) transgenic mice were exposed to Dox for 5 d. (A–B) Immunostaining for Sca-1 was performed on lung sections. Increased Sca-1 staining was detected in the peripheral lung and near the bronchoalveolar duct junctions after expression of Sox17 (arrowheads and inset). (C–E) Dual-label immunofluorescence for Sox17 (C) and Sca-1 (D) demonstrated that the Sca-1 positive cells coexpressed Sox17 (arrows). (F–N) Triple-label immunofluorescent staining was performed for CCSP, proSP-C, and Sca-1. Sca-1 colocalized with CCSP-expressing cells near the bronchoalveolar duct junctions (open arrowhead; magnified in G–J). Sca-1 positive cells located in the peripheral lung (arrowheads; magnified in K–N) did not coexpress proSP-C (arrow; F,L, and N). Scale bars, 50 µm (A–B; F), 20 µm (C–E).

    Journal: PLoS ONE

    Article Title: Sox17 Promotes Cell Cycle Progression and Inhibits TGF-?/Smad3 Signaling to Initiate Progenitor Cell Behavior in the Respiratory Epithelium

    doi: 10.1371/journal.pone.0005711

    Figure Lengend Snippet: Sox17 increases Sca-1 expression adult respiratory epithelial cells. Adult CCSPrtTA control (A) and CCSPrtTA/tetO-Sox17 (B–N) transgenic mice were exposed to Dox for 5 d. (A–B) Immunostaining for Sca-1 was performed on lung sections. Increased Sca-1 staining was detected in the peripheral lung and near the bronchoalveolar duct junctions after expression of Sox17 (arrowheads and inset). (C–E) Dual-label immunofluorescence for Sox17 (C) and Sca-1 (D) demonstrated that the Sca-1 positive cells coexpressed Sox17 (arrows). (F–N) Triple-label immunofluorescent staining was performed for CCSP, proSP-C, and Sca-1. Sca-1 colocalized with CCSP-expressing cells near the bronchoalveolar duct junctions (open arrowhead; magnified in G–J). Sca-1 positive cells located in the peripheral lung (arrowheads; magnified in K–N) did not coexpress proSP-C (arrow; F,L, and N). Scale bars, 50 µm (A–B; F), 20 µm (C–E).

    Article Snippet: Immunohistochemistry was performed using primary antibodies for guinea pig-anti Sox17 (1∶10,000), rabbit anti-phospho-histone H3 (1∶1000; Santa Cruz), rabbit anti-CCSP (1∶5000), rabbit anti-Foxj1 (1∶14,000), rabbit anti-cyclin D1 (1∶400; Abcam), and rat anti-Sca-1 (1∶250; BD Pharmingen).

    Techniques: Expressing, Transgenic Assay, Mouse Assay, Immunostaining, Staining, Immunofluorescence