primary mouse osteocytes Search Results


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  • 99
    Vector Laboratories abc ap kit
    Abc Ap Kit, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 154 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC raw264 7 macrophage cell line
    Effects of COX-2 inhibition on MLO-Y4 cell responses to MSU crystal-stimulated <t>RAW264.7</t> macrophage conditioned medium. RAW264.7 macrophages were cultured with or without 0.5 mg/mL monosodium urate (MSU) crystals for 24 h for preparation of MSU crystal-stimulated conditioned medium and control conditioned medium, respectively. A cyclooxygenase-2 (COX-2)-specific inhibitor (SC-236) was added to MLO-Y4 cells for 1 h prior to the addition of 40% conditioned medium for 24 h. MLO-Y4 cells were then harvested for mRNA gene expression analysis and supernatants harvested for protein quantification. a Changes in mRNA expression of inflammatory genes: tumor necrosis factor (TNF)-α, COX-2, interleukin (IL)-6, and IL-11; and bone-related genes: receptor activator of nuclear factor kappa-B ligand (RANKL) and osteoprotegerin (OPG). b Changes in TNF-α, prostaglandin E 2 (PGE 2 ), and IL-6 protein levels in MLO-Y4 cell supernatants. Data shown are pooled from five biological repeats and are presented as (SEM); one-way analysis of variance (ANOVA) with post-hoc Sidak’s test between groups as indicated. NS no significant difference
    Raw264 7 Macrophage Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 124 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore anti connexin 43 antibody
    Effects of COX-2 inhibition on MLO-Y4 cell responses to MSU crystal-stimulated <t>RAW264.7</t> macrophage conditioned medium. RAW264.7 macrophages were cultured with or without 0.5 mg/mL monosodium urate (MSU) crystals for 24 h for preparation of MSU crystal-stimulated conditioned medium and control conditioned medium, respectively. A cyclooxygenase-2 (COX-2)-specific inhibitor (SC-236) was added to MLO-Y4 cells for 1 h prior to the addition of 40% conditioned medium for 24 h. MLO-Y4 cells were then harvested for mRNA gene expression analysis and supernatants harvested for protein quantification. a Changes in mRNA expression of inflammatory genes: tumor necrosis factor (TNF)-α, COX-2, interleukin (IL)-6, and IL-11; and bone-related genes: receptor activator of nuclear factor kappa-B ligand (RANKL) and osteoprotegerin (OPG). b Changes in TNF-α, prostaglandin E 2 (PGE 2 ), and IL-6 protein levels in MLO-Y4 cell supernatants. Data shown are pooled from five biological repeats and are presented as (SEM); one-way analysis of variance (ANOVA) with post-hoc Sidak’s test between groups as indicated. NS no significant difference
    Anti Connexin 43 Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1449 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore monoclonal anti vimentin antibody
    PDAC cells demonstrate different invasive phenotypes. The 3D tumor-tissue invasion model was created using 200 Pa Oligomer for both the tumor and surrounding tissue compartments. Tumor compartments were prepared with 1 × 10 7 Panc-1 or BxPC-3 cells/mL in Oligomer and cultured 5 days. Constructs were fixed, cryosectioned, and immunostained for ( a ) <t>vimentin</t> (red) and E-cadherin (yellow) with nuclear counterstaining (blue; Hoechst 33342). Images represent maximum projections of 20 μm confocal z-stacks. ( b ) Sections were stained for vimentin (red) and imaged with confocal reflection microscopy to visualize matrix microstructure. Images represent maximum projections of 10 μm confocal z-stacks. Yellow arrowheads denote matrix remodeling and alignment. Scale bars = 50 μm.
    Monoclonal Anti Vimentin Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1840 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ucp1  (Abcam)
    94
    Abcam ucp1
    Increased sclerostin secretion from Gsα-deficient osteocytes contributes to beige adipogenesis. <t>UCP1</t> and PGC1α expressions in SVF from wild-type InWAT undergoing beige adipogenesis in response to ( A ) varying doses of sclerostin ( n = 3), and ( B ) Wnt3a and/or sclerostin treatment ( n = 3). ( C ) GWAT organ weight and ( D ) UCP1 gene expression in GWAT in wild-type mice treated with 100 μg/kg sclerostin every day for 3 weeks. ( E ) Sclerostin secretion into CM in CRISPR-mediated Gsα knockout cells and after sclerostin depletion using antibody ( n = 3). ( F ) UCP1, PGC1α, and Elovl3 expression in primary SVF after treatment with CM from Ocy454 cells. Sclerostin-neutralizing antibody treatment of control and DMP1-GsαKO mice for 2 weeks showing ( G ) serum sclerostin changes and ( H ) gene expression changes in InWAT tissues ( n = 4, each group). All data: mean ± SE; * p
    Ucp1, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 1844 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc cleaved caspase 3
    Accelerated osteoblast apoptosis and suppressed osteoblast proliferation following SCI. Sections through the distal femoral metaphysis were stained with cleaved <t>caspase-3</t> (CC3) antibody to identify apoptotic osteoblasts in uninjured rats ( top , panel
    Cleaved Caspase 3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 38170 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher alpha mem
    Accelerated osteoblast apoptosis and suppressed osteoblast proliferation following SCI. Sections through the distal femoral metaphysis were stained with cleaved <t>caspase-3</t> (CC3) antibody to identify apoptotic osteoblasts in uninjured rats ( top , panel
    Alpha Mem, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1522 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher alamarblue reagent
    The direct effects of MSU crystals on osteocyte viability. The <t>alamarBlue®</t> assay was used to determine the viability of a MLO-Y4 cells and primary mouse osteocytes cultured with monosodium urate (MSU) crystals for 24 h, b MLO-Y4 cells cultured with soluble urate for 24 h, and c MLO-Y4 cells cultured with different types of crystals for 24 h. Viability was assessed 24 and 48 h after the addition of crystals or soluble urate. Data shown are pooled from three to four biological repeats and are presented as mean (SEM); by two-way ANOVA a P Interaction
    Alamarblue Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1074 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher fetal bovine serum
    The direct effects of MSU crystals on osteocyte viability. The <t>alamarBlue®</t> assay was used to determine the viability of a MLO-Y4 cells and primary mouse osteocytes cultured with monosodium urate (MSU) crystals for 24 h, b MLO-Y4 cells cultured with soluble urate for 24 h, and c MLO-Y4 cells cultured with different types of crystals for 24 h. Viability was assessed 24 and 48 h after the addition of crystals or soluble urate. Data shown are pooled from three to four biological repeats and are presented as mean (SEM); by two-way ANOVA a P Interaction
    Fetal Bovine Serum, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 225914 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher α minimal essential medium α mem
    The direct effects of MSU crystals on osteocyte viability. The <t>alamarBlue®</t> assay was used to determine the viability of a MLO-Y4 cells and primary mouse osteocytes cultured with monosodium urate (MSU) crystals for 24 h, b MLO-Y4 cells cultured with soluble urate for 24 h, and c MLO-Y4 cells cultured with different types of crystals for 24 h. Viability was assessed 24 and 48 h after the addition of crystals or soluble urate. Data shown are pooled from three to four biological repeats and are presented as mean (SEM); by two-way ANOVA a P Interaction
    α Minimal Essential Medium α Mem, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 314 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Vector Biolabs cuedc2
    <t>CUEDC2</t> inhibits STAT3-mediated osteogenesis by regulating SOCS3 protein stability. a – c The level of SOCS1, SOCS3, and phosphorylated STAT3 proteins was evaluated by western blotting. The relative protein levels of the indicated proteins were calculated after normalization to β-ACTIN. * P
    Cuedc2, supplied by Vector Biolabs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Novartis imatinib mesylate
    <t>Imatinib</t> <t>mesylate</t> inhibits in a dose dependent manner the osteosarcoma cells proliferation. Human (HOS, MG63) ( A ) , mouse ( POS-1, MOS-J ) ( B ) and rat (OSRGA) ( C ) osteosarcoma cell lines were treated by increasing concentration of imatinib mesylate (0.1–40 µM) for 72 hours. The number of viable cells was then determined using an XTT assay. ( D ) Table summarizing the IC50 and IC90 of each cell lines studied. Graphs represent the average values of three independent experiments performed in triplicate.
    Imatinib Mesylate, supplied by Novartis, used in various techniques. Bioz Stars score: 92/100, based on 2328 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies collagen coated plate
    <t>Imatinib</t> <t>mesylate</t> inhibits in a dose dependent manner the osteosarcoma cells proliferation. Human (HOS, MG63) ( A ) , mouse ( POS-1, MOS-J ) ( B ) and rat (OSRGA) ( C ) osteosarcoma cell lines were treated by increasing concentration of imatinib mesylate (0.1–40 µM) for 72 hours. The number of viable cells was then determined using an XTT assay. ( D ) Table summarizing the IC50 and IC90 of each cell lines studied. Graphs represent the average values of three independent experiments performed in triplicate.
    Collagen Coated Plate, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore l glutamine
    <t>Imatinib</t> <t>mesylate</t> inhibits in a dose dependent manner the osteosarcoma cells proliferation. Human (HOS, MG63) ( A ) , mouse ( POS-1, MOS-J ) ( B ) and rat (OSRGA) ( C ) osteosarcoma cell lines were treated by increasing concentration of imatinib mesylate (0.1–40 µM) for 72 hours. The number of viable cells was then determined using an XTT assay. ( D ) Table summarizing the IC50 and IC90 of each cell lines studied. Graphs represent the average values of three independent experiments performed in triplicate.
    L Glutamine, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 52942 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies respiration analyzer
    <t>Imatinib</t> <t>mesylate</t> inhibits in a dose dependent manner the osteosarcoma cells proliferation. Human (HOS, MG63) ( A ) , mouse ( POS-1, MOS-J ) ( B ) and rat (OSRGA) ( C ) osteosarcoma cell lines were treated by increasing concentration of imatinib mesylate (0.1–40 µM) for 72 hours. The number of viable cells was then determined using an XTT assay. ( D ) Table summarizing the IC50 and IC90 of each cell lines studied. Graphs represent the average values of three independent experiments performed in triplicate.
    Respiration Analyzer, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore collagenase type ia
    <t>Imatinib</t> <t>mesylate</t> inhibits in a dose dependent manner the osteosarcoma cells proliferation. Human (HOS, MG63) ( A ) , mouse ( POS-1, MOS-J ) ( B ) and rat (OSRGA) ( C ) osteosarcoma cell lines were treated by increasing concentration of imatinib mesylate (0.1–40 µM) for 72 hours. The number of viable cells was then determined using an XTT assay. ( D ) Table summarizing the IC50 and IC90 of each cell lines studied. Graphs represent the average values of three independent experiments performed in triplicate.
    Collagenase Type Ia, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1992 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore collagenase
    <t>Imatinib</t> <t>mesylate</t> inhibits in a dose dependent manner the osteosarcoma cells proliferation. Human (HOS, MG63) ( A ) , mouse ( POS-1, MOS-J ) ( B ) and rat (OSRGA) ( C ) osteosarcoma cell lines were treated by increasing concentration of imatinib mesylate (0.1–40 µM) for 72 hours. The number of viable cells was then determined using an XTT assay. ( D ) Table summarizing the IC50 and IC90 of each cell lines studied. Graphs represent the average values of three independent experiments performed in triplicate.
    Collagenase, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 20723 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies mouse anti human cd31 primary antibody
    Deletion of PHD2 in osteocytes stimulates angiogenesis. a , b <t>CD31</t> immunostaining ( a ) of the tibial metaphysis with quantification ( b ) of blood vessel number and size in 8-week-old mice ( n = 8 Phd2 ot+ –10 Phd2 ot − ). c , d CD31 immunostaining ( c ) of the cortical diaphysis of tibiae with quantification ( d ) of blood vessel number and size ( n = 8 Phd2 ot+ –10 Phd2 ot− ). e Vegf and Plgf mRNA levels in femora of 8-week-old mice ( n = 8 Phd2 ot+ –10 Phd2 ot− ) and in vehicle (IDG VEH ) or IOX2-treated IDG-SW3 (IDG IOX2 ) cells ( n = 3). f Scheme of experimental design. HIF-1α (shH1) or HIF-2α (shH2) were silenced in IDG IOX2 cells and a scrambled shRNA (shScr) was used as control. Conditioned medium was subsequently used to culture HUVECs in monoculture or in co-culture with IDG-SW3 cells. g Proliferation of HUVECs in monoculture according to the scheme in f ( n = 3). h , i Human CD31 (hCD31) immunostaining ( h ) and quantification ( i ) of hCD31-positive surface in HUVECs co-cultured with IDG-SW3 cells, according to the scheme in f . When indicated, cells were treated with anti-VEGF 164 antibody or recombinant mouse soluble VEGFR-1 (sFlt) ( n = 3). Data are means ± SEM. * p
    Mouse Anti Human Cd31 Primary Antibody, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 91/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Corning Life Sciences rat collagen type i
    Deletion of PHD2 in osteocytes stimulates angiogenesis. a , b <t>CD31</t> immunostaining ( a ) of the tibial metaphysis with quantification ( b ) of blood vessel number and size in 8-week-old mice ( n = 8 Phd2 ot+ –10 Phd2 ot − ). c , d CD31 immunostaining ( c ) of the cortical diaphysis of tibiae with quantification ( d ) of blood vessel number and size ( n = 8 Phd2 ot+ –10 Phd2 ot− ). e Vegf and Plgf mRNA levels in femora of 8-week-old mice ( n = 8 Phd2 ot+ –10 Phd2 ot− ) and in vehicle (IDG VEH ) or IOX2-treated IDG-SW3 (IDG IOX2 ) cells ( n = 3). f Scheme of experimental design. HIF-1α (shH1) or HIF-2α (shH2) were silenced in IDG IOX2 cells and a scrambled shRNA (shScr) was used as control. Conditioned medium was subsequently used to culture HUVECs in monoculture or in co-culture with IDG-SW3 cells. g Proliferation of HUVECs in monoculture according to the scheme in f ( n = 3). h , i Human CD31 (hCD31) immunostaining ( h ) and quantification ( i ) of hCD31-positive surface in HUVECs co-cultured with IDG-SW3 cells, according to the scheme in f . When indicated, cells were treated with anti-VEGF 164 antibody or recombinant mouse soluble VEGFR-1 (sFlt) ( n = 3). Data are means ± SEM. * p
    Rat Collagen Type I, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 93/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore iscove s modified dulbecco s medium
    Deletion of PHD2 in osteocytes stimulates angiogenesis. a , b <t>CD31</t> immunostaining ( a ) of the tibial metaphysis with quantification ( b ) of blood vessel number and size in 8-week-old mice ( n = 8 Phd2 ot+ –10 Phd2 ot − ). c , d CD31 immunostaining ( c ) of the cortical diaphysis of tibiae with quantification ( d ) of blood vessel number and size ( n = 8 Phd2 ot+ –10 Phd2 ot− ). e Vegf and Plgf mRNA levels in femora of 8-week-old mice ( n = 8 Phd2 ot+ –10 Phd2 ot− ) and in vehicle (IDG VEH ) or IOX2-treated IDG-SW3 (IDG IOX2 ) cells ( n = 3). f Scheme of experimental design. HIF-1α (shH1) or HIF-2α (shH2) were silenced in IDG IOX2 cells and a scrambled shRNA (shScr) was used as control. Conditioned medium was subsequently used to culture HUVECs in monoculture or in co-culture with IDG-SW3 cells. g Proliferation of HUVECs in monoculture according to the scheme in f ( n = 3). h , i Human CD31 (hCD31) immunostaining ( h ) and quantification ( i ) of hCD31-positive surface in HUVECs co-cultured with IDG-SW3 cells, according to the scheme in f . When indicated, cells were treated with anti-VEGF 164 antibody or recombinant mouse soluble VEGFR-1 (sFlt) ( n = 3). Data are means ± SEM. * p
    Iscove S Modified Dulbecco S Medium, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1773 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen rneasy kit
    Deletion of PHD2 in osteocytes stimulates angiogenesis. a , b <t>CD31</t> immunostaining ( a ) of the tibial metaphysis with quantification ( b ) of blood vessel number and size in 8-week-old mice ( n = 8 Phd2 ot+ –10 Phd2 ot − ). c , d CD31 immunostaining ( c ) of the cortical diaphysis of tibiae with quantification ( d ) of blood vessel number and size ( n = 8 Phd2 ot+ –10 Phd2 ot− ). e Vegf and Plgf mRNA levels in femora of 8-week-old mice ( n = 8 Phd2 ot+ –10 Phd2 ot− ) and in vehicle (IDG VEH ) or IOX2-treated IDG-SW3 (IDG IOX2 ) cells ( n = 3). f Scheme of experimental design. HIF-1α (shH1) or HIF-2α (shH2) were silenced in IDG IOX2 cells and a scrambled shRNA (shScr) was used as control. Conditioned medium was subsequently used to culture HUVECs in monoculture or in co-culture with IDG-SW3 cells. g Proliferation of HUVECs in monoculture according to the scheme in f ( n = 3). h , i Human CD31 (hCD31) immunostaining ( h ) and quantification ( i ) of hCD31-positive surface in HUVECs co-cultured with IDG-SW3 cells, according to the scheme in f . When indicated, cells were treated with anti-VEGF 164 antibody or recombinant mouse soluble VEGFR-1 (sFlt) ( n = 3). Data are means ± SEM. * p
    Rneasy Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 110557 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer tsa cyanine 3 system
    Deletion of PHD2 in osteocytes stimulates angiogenesis. a , b <t>CD31</t> immunostaining ( a ) of the tibial metaphysis with quantification ( b ) of blood vessel number and size in 8-week-old mice ( n = 8 Phd2 ot+ –10 Phd2 ot − ). c , d CD31 immunostaining ( c ) of the cortical diaphysis of tibiae with quantification ( d ) of blood vessel number and size ( n = 8 Phd2 ot+ –10 Phd2 ot− ). e Vegf and Plgf mRNA levels in femora of 8-week-old mice ( n = 8 Phd2 ot+ –10 Phd2 ot− ) and in vehicle (IDG VEH ) or IOX2-treated IDG-SW3 (IDG IOX2 ) cells ( n = 3). f Scheme of experimental design. HIF-1α (shH1) or HIF-2α (shH2) were silenced in IDG IOX2 cells and a scrambled shRNA (shScr) was used as control. Conditioned medium was subsequently used to culture HUVECs in monoculture or in co-culture with IDG-SW3 cells. g Proliferation of HUVECs in monoculture according to the scheme in f ( n = 3). h , i Human CD31 (hCD31) immunostaining ( h ) and quantification ( i ) of hCD31-positive surface in HUVECs co-cultured with IDG-SW3 cells, according to the scheme in f . When indicated, cells were treated with anti-VEGF 164 antibody or recombinant mouse soluble VEGFR-1 (sFlt) ( n = 3). Data are means ± SEM. * p
    Tsa Cyanine 3 System, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 99/100, based on 338 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC umr 106 cells
    FGF23 stimulation by hypoxia in vitro and in vivo. ( A ) FGF23 mRNA was significantly elevated in <t>UMR-106</t> cells cultured under hypoxic conditions for 24 to 48 hours. In parallel, immunoblots (chart inset) for HIF1α (upper panel) showed an increase
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    Thermo Fisher 1x antibiotic antimycotic
    FGF23 stimulation by hypoxia in vitro and in vivo. ( A ) FGF23 mRNA was significantly elevated in <t>UMR-106</t> cells cultured under hypoxic conditions for 24 to 48 hours. In parallel, immunoblots (chart inset) for HIF1α (upper panel) showed an increase
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    Thermo Fisher alamarblue assay
    FGF23 stimulation by hypoxia in vitro and in vivo. ( A ) FGF23 mRNA was significantly elevated in <t>UMR-106</t> cells cultured under hypoxic conditions for 24 to 48 hours. In parallel, immunoblots (chart inset) for HIF1α (upper panel) showed an increase
    Alamarblue Assay, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1829 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Effects of COX-2 inhibition on MLO-Y4 cell responses to MSU crystal-stimulated RAW264.7 macrophage conditioned medium. RAW264.7 macrophages were cultured with or without 0.5 mg/mL monosodium urate (MSU) crystals for 24 h for preparation of MSU crystal-stimulated conditioned medium and control conditioned medium, respectively. A cyclooxygenase-2 (COX-2)-specific inhibitor (SC-236) was added to MLO-Y4 cells for 1 h prior to the addition of 40% conditioned medium for 24 h. MLO-Y4 cells were then harvested for mRNA gene expression analysis and supernatants harvested for protein quantification. a Changes in mRNA expression of inflammatory genes: tumor necrosis factor (TNF)-α, COX-2, interleukin (IL)-6, and IL-11; and bone-related genes: receptor activator of nuclear factor kappa-B ligand (RANKL) and osteoprotegerin (OPG). b Changes in TNF-α, prostaglandin E 2 (PGE 2 ), and IL-6 protein levels in MLO-Y4 cell supernatants. Data shown are pooled from five biological repeats and are presented as (SEM); one-way analysis of variance (ANOVA) with post-hoc Sidak’s test between groups as indicated. NS no significant difference

    Journal: Arthritis Research & Therapy

    Article Title: Monosodium urate crystals reduce osteocyte viability and indirectly promote a shift in osteocyte function towards a proinflammatory and proresorptive state

    doi: 10.1186/s13075-018-1704-y

    Figure Lengend Snippet: Effects of COX-2 inhibition on MLO-Y4 cell responses to MSU crystal-stimulated RAW264.7 macrophage conditioned medium. RAW264.7 macrophages were cultured with or without 0.5 mg/mL monosodium urate (MSU) crystals for 24 h for preparation of MSU crystal-stimulated conditioned medium and control conditioned medium, respectively. A cyclooxygenase-2 (COX-2)-specific inhibitor (SC-236) was added to MLO-Y4 cells for 1 h prior to the addition of 40% conditioned medium for 24 h. MLO-Y4 cells were then harvested for mRNA gene expression analysis and supernatants harvested for protein quantification. a Changes in mRNA expression of inflammatory genes: tumor necrosis factor (TNF)-α, COX-2, interleukin (IL)-6, and IL-11; and bone-related genes: receptor activator of nuclear factor kappa-B ligand (RANKL) and osteoprotegerin (OPG). b Changes in TNF-α, prostaglandin E 2 (PGE 2 ), and IL-6 protein levels in MLO-Y4 cell supernatants. Data shown are pooled from five biological repeats and are presented as (SEM); one-way analysis of variance (ANOVA) with post-hoc Sidak’s test between groups as indicated. NS no significant difference

    Article Snippet: The RAW264.7 macrophage cell line (ATCC, Manassas, USA) was maintained in Iscove’s modified Dulbecco’s medium supplemented with 1 mM l -glutamine (both from Sigma-Aldrich) and 10% FBS.

    Techniques: Inhibition, Cell Culture, Expressing

    Secretion of proinflammatory mediators by MLO-Y4 cells in response to MSU crystal-stimulated RAW264.7 macrophages. RAW264.7 macrophages were cultured with or without 0.5 mg/mL monosodium urate (MSU) crystals for 24 h for preparation of MSU crystal-stimulated conditioned medium and control conditioned medium, respectively. Conditioned medium preparations were added to MLO-Y4 cells (40% final concentration in a well) for 24 h and supernatants harvested. The concentrations of a tumor necrosis factor (TNF)-α, b prostaglandin E 2 (PGE 2 ), c interleukin (IL)-6, and d osteoprotegerin (OPG) protein in the RAW264.7 macrophage conditioned medium samples (control and MSU crystal-stimulated) and the MLO-Y4 cell supernatants were measured by ELISA. Data shown are pooled from three biological repeats and are presented as mean (SEM); one-way analysis of variance (ANOVA) with post-hoc Sidak’s test between groups as indicated. NS no significant difference

    Journal: Arthritis Research & Therapy

    Article Title: Monosodium urate crystals reduce osteocyte viability and indirectly promote a shift in osteocyte function towards a proinflammatory and proresorptive state

    doi: 10.1186/s13075-018-1704-y

    Figure Lengend Snippet: Secretion of proinflammatory mediators by MLO-Y4 cells in response to MSU crystal-stimulated RAW264.7 macrophages. RAW264.7 macrophages were cultured with or without 0.5 mg/mL monosodium urate (MSU) crystals for 24 h for preparation of MSU crystal-stimulated conditioned medium and control conditioned medium, respectively. Conditioned medium preparations were added to MLO-Y4 cells (40% final concentration in a well) for 24 h and supernatants harvested. The concentrations of a tumor necrosis factor (TNF)-α, b prostaglandin E 2 (PGE 2 ), c interleukin (IL)-6, and d osteoprotegerin (OPG) protein in the RAW264.7 macrophage conditioned medium samples (control and MSU crystal-stimulated) and the MLO-Y4 cell supernatants were measured by ELISA. Data shown are pooled from three biological repeats and are presented as mean (SEM); one-way analysis of variance (ANOVA) with post-hoc Sidak’s test between groups as indicated. NS no significant difference

    Article Snippet: The RAW264.7 macrophage cell line (ATCC, Manassas, USA) was maintained in Iscove’s modified Dulbecco’s medium supplemented with 1 mM l -glutamine (both from Sigma-Aldrich) and 10% FBS.

    Techniques: Cell Culture, Concentration Assay, Enzyme-linked Immunosorbent Assay

    Indirect effects of MSU crystal-stimulated RAW264.7 macrophage conditioned medium on MLO-Y4 cell gene expression. RAW264.7 macrophages were cultured with or without 0.5 mg/mL monosodium urate (MSU) crystals for 24 h for preparation of MSU crystal-stimulated conditioned medium and control conditioned medium, respectively. Conditioned medium preparations were added to MLO-Y4 cells (40% final concentration in a well) for 0, 1, 6, and 24 h. MLO-Y4 cells were harvested and real-time PCR was used to determine changes in the relative mRNA expression levels of a bone-related and b inflammatory genes. Data shown are pooled from three biological repeats and are presented as mean (SEM); two-way ANOVA P Interaction = 0.007 for Tnfrsf11b , P Interaction = 0.0005 for Tnfa , P Interaction

    Journal: Arthritis Research & Therapy

    Article Title: Monosodium urate crystals reduce osteocyte viability and indirectly promote a shift in osteocyte function towards a proinflammatory and proresorptive state

    doi: 10.1186/s13075-018-1704-y

    Figure Lengend Snippet: Indirect effects of MSU crystal-stimulated RAW264.7 macrophage conditioned medium on MLO-Y4 cell gene expression. RAW264.7 macrophages were cultured with or without 0.5 mg/mL monosodium urate (MSU) crystals for 24 h for preparation of MSU crystal-stimulated conditioned medium and control conditioned medium, respectively. Conditioned medium preparations were added to MLO-Y4 cells (40% final concentration in a well) for 0, 1, 6, and 24 h. MLO-Y4 cells were harvested and real-time PCR was used to determine changes in the relative mRNA expression levels of a bone-related and b inflammatory genes. Data shown are pooled from three biological repeats and are presented as mean (SEM); two-way ANOVA P Interaction = 0.007 for Tnfrsf11b , P Interaction = 0.0005 for Tnfa , P Interaction

    Article Snippet: The RAW264.7 macrophage cell line (ATCC, Manassas, USA) was maintained in Iscove’s modified Dulbecco’s medium supplemented with 1 mM l -glutamine (both from Sigma-Aldrich) and 10% FBS.

    Techniques: Expressing, Cell Culture, Concentration Assay, Real-time Polymerase Chain Reaction

    PDAC cells demonstrate different invasive phenotypes. The 3D tumor-tissue invasion model was created using 200 Pa Oligomer for both the tumor and surrounding tissue compartments. Tumor compartments were prepared with 1 × 10 7 Panc-1 or BxPC-3 cells/mL in Oligomer and cultured 5 days. Constructs were fixed, cryosectioned, and immunostained for ( a ) vimentin (red) and E-cadherin (yellow) with nuclear counterstaining (blue; Hoechst 33342). Images represent maximum projections of 20 μm confocal z-stacks. ( b ) Sections were stained for vimentin (red) and imaged with confocal reflection microscopy to visualize matrix microstructure. Images represent maximum projections of 10 μm confocal z-stacks. Yellow arrowheads denote matrix remodeling and alignment. Scale bars = 50 μm.

    Journal: Scientific Reports

    Article Title: Development of a Novel 3D Tumor-tissue Invasion Model for High-throughput, High-content Phenotypic Drug Screening

    doi: 10.1038/s41598-018-31138-6

    Figure Lengend Snippet: PDAC cells demonstrate different invasive phenotypes. The 3D tumor-tissue invasion model was created using 200 Pa Oligomer for both the tumor and surrounding tissue compartments. Tumor compartments were prepared with 1 × 10 7 Panc-1 or BxPC-3 cells/mL in Oligomer and cultured 5 days. Constructs were fixed, cryosectioned, and immunostained for ( a ) vimentin (red) and E-cadherin (yellow) with nuclear counterstaining (blue; Hoechst 33342). Images represent maximum projections of 20 μm confocal z-stacks. ( b ) Sections were stained for vimentin (red) and imaged with confocal reflection microscopy to visualize matrix microstructure. Images represent maximum projections of 10 μm confocal z-stacks. Yellow arrowheads denote matrix remodeling and alignment. Scale bars = 50 μm.

    Article Snippet: Primary antibodies included mouse anti-vimentin (V6389, Sigma Aldrich) and rabbit anti-E-cadherin (24E10, Cell Signaling Technologies, Danvers, MA).

    Techniques: Cell Culture, Construct, Staining, Microscopy

    CAFs enhance invasiveness of patient-derived PDAC cells. The 3D tumor-tissue invasion model was prepared with 200 Pa Oligomer for both the tumor and surrounding tissue compartments. Tumor compartments were created with 10.05 alone or 10.05 + CAFS (at 1:1 ratio) at 1 × 10 7 cells/mL in Oligomer and cultured 4 days. ( a ) Images represent nine fields of view, each of which is a maximum projection of a 400 μm confocal z-stack. Red = tumor cells (TdT) and green = CAFs (EGFP). Scale bars = 200 μm. ( b ) Images represent maximum projections of 10 μm confocal z-stacks from cryosectioned constructs. Confocal reflection microscopy (white) was used to visualize matrix microstructure. Yellow arrowheads denote matrix alignment and remodeling; scale bars = 20 μm. ( c ) Images represent maximum projections of a 20 μm confocal z-stacks of cryosectioned constructs stained for E-cadherin (yellow) and vimentin (blue). Final panel represents a 3X zoom of boxed region in overlay panel. White arrowhead denotes direct interaction between tumor cell and CAF; scale bars = 50 μm.

    Journal: Scientific Reports

    Article Title: Development of a Novel 3D Tumor-tissue Invasion Model for High-throughput, High-content Phenotypic Drug Screening

    doi: 10.1038/s41598-018-31138-6

    Figure Lengend Snippet: CAFs enhance invasiveness of patient-derived PDAC cells. The 3D tumor-tissue invasion model was prepared with 200 Pa Oligomer for both the tumor and surrounding tissue compartments. Tumor compartments were created with 10.05 alone or 10.05 + CAFS (at 1:1 ratio) at 1 × 10 7 cells/mL in Oligomer and cultured 4 days. ( a ) Images represent nine fields of view, each of which is a maximum projection of a 400 μm confocal z-stack. Red = tumor cells (TdT) and green = CAFs (EGFP). Scale bars = 200 μm. ( b ) Images represent maximum projections of 10 μm confocal z-stacks from cryosectioned constructs. Confocal reflection microscopy (white) was used to visualize matrix microstructure. Yellow arrowheads denote matrix alignment and remodeling; scale bars = 20 μm. ( c ) Images represent maximum projections of a 20 μm confocal z-stacks of cryosectioned constructs stained for E-cadherin (yellow) and vimentin (blue). Final panel represents a 3X zoom of boxed region in overlay panel. White arrowhead denotes direct interaction between tumor cell and CAF; scale bars = 50 μm.

    Article Snippet: Primary antibodies included mouse anti-vimentin (V6389, Sigma Aldrich) and rabbit anti-E-cadherin (24E10, Cell Signaling Technologies, Danvers, MA).

    Techniques: Derivative Assay, Cell Culture, Construct, Microscopy, Staining

    Increased sclerostin secretion from Gsα-deficient osteocytes contributes to beige adipogenesis. UCP1 and PGC1α expressions in SVF from wild-type InWAT undergoing beige adipogenesis in response to ( A ) varying doses of sclerostin ( n = 3), and ( B ) Wnt3a and/or sclerostin treatment ( n = 3). ( C ) GWAT organ weight and ( D ) UCP1 gene expression in GWAT in wild-type mice treated with 100 μg/kg sclerostin every day for 3 weeks. ( E ) Sclerostin secretion into CM in CRISPR-mediated Gsα knockout cells and after sclerostin depletion using antibody ( n = 3). ( F ) UCP1, PGC1α, and Elovl3 expression in primary SVF after treatment with CM from Ocy454 cells. Sclerostin-neutralizing antibody treatment of control and DMP1-GsαKO mice for 2 weeks showing ( G ) serum sclerostin changes and ( H ) gene expression changes in InWAT tissues ( n = 4, each group). All data: mean ± SE; * p

    Journal: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

    Article Title: Osteocyte-Secreted Wnt Signaling Inhibitor Sclerostin Contributes to Beige Adipogenesis in Peripheral Fat Depots

    doi: 10.1002/jbmr.3001

    Figure Lengend Snippet: Increased sclerostin secretion from Gsα-deficient osteocytes contributes to beige adipogenesis. UCP1 and PGC1α expressions in SVF from wild-type InWAT undergoing beige adipogenesis in response to ( A ) varying doses of sclerostin ( n = 3), and ( B ) Wnt3a and/or sclerostin treatment ( n = 3). ( C ) GWAT organ weight and ( D ) UCP1 gene expression in GWAT in wild-type mice treated with 100 μg/kg sclerostin every day for 3 weeks. ( E ) Sclerostin secretion into CM in CRISPR-mediated Gsα knockout cells and after sclerostin depletion using antibody ( n = 3). ( F ) UCP1, PGC1α, and Elovl3 expression in primary SVF after treatment with CM from Ocy454 cells. Sclerostin-neutralizing antibody treatment of control and DMP1-GsαKO mice for 2 weeks showing ( G ) serum sclerostin changes and ( H ) gene expression changes in InWAT tissues ( n = 4, each group). All data: mean ± SE; * p

    Article Snippet: Treatment with 20% (vol/vol) CM from Ocy454 cells significantly increased expression of UCP1 and PGC1α in primary adipocytes undergoing beige adipogenic differentiation compared to no treatment control group ( ).

    Techniques: Expressing, Mouse Assay, CRISPR, Knock-Out

    Beige adipogenesis is increased in mice lacking Gsα in osteolineage cells. Representative H E staining of GWATs from ( A ) DMP1-GsαKO, ( B ) OC-GsαKO, and ( C ) DMP1-ERT2-GsαKO mice with corresponding control littermates ( n = 4). UCP1 IHC staining from ( D ) DMP1-GsαKO, ( E ) OC-GsαKO, and ( F ) DMP1-ERT2-GsαKO mice ( n = 3). Gene expressions of beige adipogenesis markers PRDM16, Cox5b, Dio2, Cidea, Elovl3, and UCP1 in InWAT from ( G ) DMP1-GsαKO, ( H ) OC-GsαKO, and ( I ) DMP1-ERT2-GsαKO mice with corresponding control littermates ( n = 5). Expressions of white adipogenesis markers PPARg, CEBPα, SREBP1, PPARα, PLIN, Acox1, and Acls1 in InWAT of ( J ) DMP1-GsαKO, ( K ) OC-GsαKO, and ( L ) DMP1-ERT2-GsαKO mice with corresponding control littermates ( n = 5). All data: mean ± SE; * p

    Journal: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

    Article Title: Osteocyte-Secreted Wnt Signaling Inhibitor Sclerostin Contributes to Beige Adipogenesis in Peripheral Fat Depots

    doi: 10.1002/jbmr.3001

    Figure Lengend Snippet: Beige adipogenesis is increased in mice lacking Gsα in osteolineage cells. Representative H E staining of GWATs from ( A ) DMP1-GsαKO, ( B ) OC-GsαKO, and ( C ) DMP1-ERT2-GsαKO mice with corresponding control littermates ( n = 4). UCP1 IHC staining from ( D ) DMP1-GsαKO, ( E ) OC-GsαKO, and ( F ) DMP1-ERT2-GsαKO mice ( n = 3). Gene expressions of beige adipogenesis markers PRDM16, Cox5b, Dio2, Cidea, Elovl3, and UCP1 in InWAT from ( G ) DMP1-GsαKO, ( H ) OC-GsαKO, and ( I ) DMP1-ERT2-GsαKO mice with corresponding control littermates ( n = 5). Expressions of white adipogenesis markers PPARg, CEBPα, SREBP1, PPARα, PLIN, Acox1, and Acls1 in InWAT of ( J ) DMP1-GsαKO, ( K ) OC-GsαKO, and ( L ) DMP1-ERT2-GsαKO mice with corresponding control littermates ( n = 5). All data: mean ± SE; * p

    Article Snippet: Treatment with 20% (vol/vol) CM from Ocy454 cells significantly increased expression of UCP1 and PGC1α in primary adipocytes undergoing beige adipogenic differentiation compared to no treatment control group ( ).

    Techniques: Mouse Assay, Staining, Immunohistochemistry

    Accelerated osteoblast apoptosis and suppressed osteoblast proliferation following SCI. Sections through the distal femoral metaphysis were stained with cleaved caspase-3 (CC3) antibody to identify apoptotic osteoblasts in uninjured rats ( top , panel

    Journal: Translational stroke research

    Article Title: Severe Spinal Cord Injury Causes Immediate Multi-cellular Dysfunction at the Chondro-Osseous Junction

    doi: 10.1007/s12975-011-0118-9

    Figure Lengend Snippet: Accelerated osteoblast apoptosis and suppressed osteoblast proliferation following SCI. Sections through the distal femoral metaphysis were stained with cleaved caspase-3 (CC3) antibody to identify apoptotic osteoblasts in uninjured rats ( top , panel

    Article Snippet: Adjacent sections were stained for cleaved Caspase-3 (Cell Signaling Technology #9661, dilution 1:800) using the ABC method and visualized by light microscopy.

    Techniques: Staining

    SCI-induced apoptosis. Western blot analysis was performed on protein extracts derived from the distal femoral metaphysis of uninjured and injured rats. Cleaved caspase 3 (17/19 kDa) and cleaved caspase 7 (20 kDa) were detected in the day 3 and day 5

    Journal: Translational stroke research

    Article Title: Severe Spinal Cord Injury Causes Immediate Multi-cellular Dysfunction at the Chondro-Osseous Junction

    doi: 10.1007/s12975-011-0118-9

    Figure Lengend Snippet: SCI-induced apoptosis. Western blot analysis was performed on protein extracts derived from the distal femoral metaphysis of uninjured and injured rats. Cleaved caspase 3 (17/19 kDa) and cleaved caspase 7 (20 kDa) were detected in the day 3 and day 5

    Article Snippet: Adjacent sections were stained for cleaved Caspase-3 (Cell Signaling Technology #9661, dilution 1:800) using the ABC method and visualized by light microscopy.

    Techniques: Western Blot, Derivative Assay

    The direct effects of MSU crystals on osteocyte viability. The alamarBlue® assay was used to determine the viability of a MLO-Y4 cells and primary mouse osteocytes cultured with monosodium urate (MSU) crystals for 24 h, b MLO-Y4 cells cultured with soluble urate for 24 h, and c MLO-Y4 cells cultured with different types of crystals for 24 h. Viability was assessed 24 and 48 h after the addition of crystals or soluble urate. Data shown are pooled from three to four biological repeats and are presented as mean (SEM); by two-way ANOVA a P Interaction

    Journal: Arthritis Research & Therapy

    Article Title: Monosodium urate crystals reduce osteocyte viability and indirectly promote a shift in osteocyte function towards a proinflammatory and proresorptive state

    doi: 10.1186/s13075-018-1704-y

    Figure Lengend Snippet: The direct effects of MSU crystals on osteocyte viability. The alamarBlue® assay was used to determine the viability of a MLO-Y4 cells and primary mouse osteocytes cultured with monosodium urate (MSU) crystals for 24 h, b MLO-Y4 cells cultured with soluble urate for 24 h, and c MLO-Y4 cells cultured with different types of crystals for 24 h. Viability was assessed 24 and 48 h after the addition of crystals or soluble urate. Data shown are pooled from three to four biological repeats and are presented as mean (SEM); by two-way ANOVA a P Interaction

    Article Snippet: alamarBlue® assay for cell viability For direct assays, various concentrations of MSU crystals were added to MLO-Y4 cells or primary mouse osteocytes for 24 h. Cells were then washed to remove MSU crystals and alamarBlue® reagent (Life Technologies) was added (5% final concentration in a well) for 6 h at 37 °C.

    Techniques: Alamar Blue Assay, Cell Culture

    CUEDC2 inhibits STAT3-mediated osteogenesis by regulating SOCS3 protein stability. a – c The level of SOCS1, SOCS3, and phosphorylated STAT3 proteins was evaluated by western blotting. The relative protein levels of the indicated proteins were calculated after normalization to β-ACTIN. * P

    Journal: Cell Death & Disease

    Article Title: CUEDC2 controls osteoblast differentiation and bone formation via SOCS3–STAT3 pathway

    doi: 10.1038/s41419-020-2562-5

    Figure Lengend Snippet: CUEDC2 inhibits STAT3-mediated osteogenesis by regulating SOCS3 protein stability. a – c The level of SOCS1, SOCS3, and phosphorylated STAT3 proteins was evaluated by western blotting. The relative protein levels of the indicated proteins were calculated after normalization to β-ACTIN. * P

    Article Snippet: Next, we tested the expression of CUEDC2 during BMP2-induced osteoblast differentiation in MC3T3-E1 cells and primary BMSCs.

    Techniques: Western Blot

    Effects of CUEDC2 on osteoclast differentiation. a – c Bone marrow macrophages were infected with pMX-IRES-EGFP (pMX-GFP) or CUEDC2 retrovirus, and the cells were then cultured with M-CSF (30 ng/ml) and RANKL (100 ng/ml) for 3 days. a Total RNA was extracted from the cultured cells, and then real-time PCR was performed using CUEDC2-, NFATc1-, and TRAP-specific primers. ** P

    Journal: Cell Death & Disease

    Article Title: CUEDC2 controls osteoblast differentiation and bone formation via SOCS3–STAT3 pathway

    doi: 10.1038/s41419-020-2562-5

    Figure Lengend Snippet: Effects of CUEDC2 on osteoclast differentiation. a – c Bone marrow macrophages were infected with pMX-IRES-EGFP (pMX-GFP) or CUEDC2 retrovirus, and the cells were then cultured with M-CSF (30 ng/ml) and RANKL (100 ng/ml) for 3 days. a Total RNA was extracted from the cultured cells, and then real-time PCR was performed using CUEDC2-, NFATc1-, and TRAP-specific primers. ** P

    Article Snippet: Next, we tested the expression of CUEDC2 during BMP2-induced osteoblast differentiation in MC3T3-E1 cells and primary BMSCs.

    Techniques: Infection, Cell Culture, Real-time Polymerase Chain Reaction

    Decreased CUEDC2 expression is correlated with bone development. a The expression levels of CUEDC2 mRNA in calvarial bone and heart tissues of mice at postnatal days 0 and 14 were evaluated by real-time PCR. CUEDC2 levels in calvarial bone at day 0 were normalized to 1, and heart tissue was used as a positive control. * P

    Journal: Cell Death & Disease

    Article Title: CUEDC2 controls osteoblast differentiation and bone formation via SOCS3–STAT3 pathway

    doi: 10.1038/s41419-020-2562-5

    Figure Lengend Snippet: Decreased CUEDC2 expression is correlated with bone development. a The expression levels of CUEDC2 mRNA in calvarial bone and heart tissues of mice at postnatal days 0 and 14 were evaluated by real-time PCR. CUEDC2 levels in calvarial bone at day 0 were normalized to 1, and heart tissue was used as a positive control. * P

    Article Snippet: Next, we tested the expression of CUEDC2 during BMP2-induced osteoblast differentiation in MC3T3-E1 cells and primary BMSCs.

    Techniques: Expressing, Mouse Assay, Real-time Polymerase Chain Reaction, Positive Control

    CUEDC2 is downregulated during osteoblast differentiation. a , b , e , f MC3T3-E1 cells and BMSCs were cultured in osteogenic medium [OM; ascorbic acid (50 µg/ml), β-glycerophosphate (5 mM), BMP2 (50 ng/ml)] for 6 days. c , d MC3T3-E1 cells were cultured with BMP2 (50, 100 ng/ml) for 2 days. a , c , e The expression of BSP, OC, and CUEDC2 mRNA was evaluated by real-time PCR using specific primers. * P

    Journal: Cell Death & Disease

    Article Title: CUEDC2 controls osteoblast differentiation and bone formation via SOCS3–STAT3 pathway

    doi: 10.1038/s41419-020-2562-5

    Figure Lengend Snippet: CUEDC2 is downregulated during osteoblast differentiation. a , b , e , f MC3T3-E1 cells and BMSCs were cultured in osteogenic medium [OM; ascorbic acid (50 µg/ml), β-glycerophosphate (5 mM), BMP2 (50 ng/ml)] for 6 days. c , d MC3T3-E1 cells were cultured with BMP2 (50, 100 ng/ml) for 2 days. a , c , e The expression of BSP, OC, and CUEDC2 mRNA was evaluated by real-time PCR using specific primers. * P

    Article Snippet: Next, we tested the expression of CUEDC2 during BMP2-induced osteoblast differentiation in MC3T3-E1 cells and primary BMSCs.

    Techniques: Cell Culture, Expressing, Real-time Polymerase Chain Reaction

    Suppression of CUEDC2 accelerates osteogenesis in vitro and in vivo. a MC3T3-E1 cells were transfected with siRNA CUEDC2 (25 nM) or siRNA control (25 nM). After 24 h, cells were cultured with GM or OM for 4 days. Real-time PCR analysis was performed using BSP-, OC-, and CUEDC2-specific primers. * P

    Journal: Cell Death & Disease

    Article Title: CUEDC2 controls osteoblast differentiation and bone formation via SOCS3–STAT3 pathway

    doi: 10.1038/s41419-020-2562-5

    Figure Lengend Snippet: Suppression of CUEDC2 accelerates osteogenesis in vitro and in vivo. a MC3T3-E1 cells were transfected with siRNA CUEDC2 (25 nM) or siRNA control (25 nM). After 24 h, cells were cultured with GM or OM for 4 days. Real-time PCR analysis was performed using BSP-, OC-, and CUEDC2-specific primers. * P

    Article Snippet: Next, we tested the expression of CUEDC2 during BMP2-induced osteoblast differentiation in MC3T3-E1 cells and primary BMSCs.

    Techniques: In Vitro, In Vivo, Transfection, Cell Culture, Real-time Polymerase Chain Reaction

    Overexpression of CUEDC2 inhibits osteogenesis in vitro and in vivo. a MC3T3-E1 cells were transfected with Myc-Flag tagged CUEDC2 (+, 500 ng; ++, 1000 ng) or an empty vector and cultured for 4 days in GM (growth medium) or OM (osteogenic medium). Real-time PCR was performed with CUEDC2-, BSP-, and OC-specific primers. *** P

    Journal: Cell Death & Disease

    Article Title: CUEDC2 controls osteoblast differentiation and bone formation via SOCS3–STAT3 pathway

    doi: 10.1038/s41419-020-2562-5

    Figure Lengend Snippet: Overexpression of CUEDC2 inhibits osteogenesis in vitro and in vivo. a MC3T3-E1 cells were transfected with Myc-Flag tagged CUEDC2 (+, 500 ng; ++, 1000 ng) or an empty vector and cultured for 4 days in GM (growth medium) or OM (osteogenic medium). Real-time PCR was performed with CUEDC2-, BSP-, and OC-specific primers. *** P

    Article Snippet: Next, we tested the expression of CUEDC2 during BMP2-induced osteoblast differentiation in MC3T3-E1 cells and primary BMSCs.

    Techniques: Over Expression, In Vitro, In Vivo, Transfection, Plasmid Preparation, Cell Culture, Real-time Polymerase Chain Reaction

    Imatinib mesylate inhibits in a dose dependent manner the osteosarcoma cells proliferation. Human (HOS, MG63) ( A ) , mouse ( POS-1, MOS-J ) ( B ) and rat (OSRGA) ( C ) osteosarcoma cell lines were treated by increasing concentration of imatinib mesylate (0.1–40 µM) for 72 hours. The number of viable cells was then determined using an XTT assay. ( D ) Table summarizing the IC50 and IC90 of each cell lines studied. Graphs represent the average values of three independent experiments performed in triplicate.

    Journal: PLoS ONE

    Article Title: Imatinib Mesylate Exerts Anti-Proliferative Effects on Osteosarcoma Cells and Inhibits the Tumour Growth in Immunocompetent Murine Models

    doi: 10.1371/journal.pone.0090795

    Figure Lengend Snippet: Imatinib mesylate inhibits in a dose dependent manner the osteosarcoma cells proliferation. Human (HOS, MG63) ( A ) , mouse ( POS-1, MOS-J ) ( B ) and rat (OSRGA) ( C ) osteosarcoma cell lines were treated by increasing concentration of imatinib mesylate (0.1–40 µM) for 72 hours. The number of viable cells was then determined using an XTT assay. ( D ) Table summarizing the IC50 and IC90 of each cell lines studied. Graphs represent the average values of three independent experiments performed in triplicate.

    Article Snippet: Imatinib mesylate (Gleevec, Novartis Pharma), a tyrosine kinase inhibitor, was originally developed for the treatment of chronic myeloid leukemia.

    Techniques: Concentration Assay, XTT Assay

    Imatinib mesylate inhibits osteosarcoma development in “preventive” and “curative” therapeutic context. Mice bearing undifferentiated POS-1 ( A, B ) or mixed osteoblastic/osteolytic MOS-J ( C–F ) osteosarcoma tumours (n = 8 per group) were assigned as control (vehicle), or imatinib mesylate (25, 50 or 100 mg/kg, daily oral administration). The treatment started 1 day after tumour cell inoculation (« preventive » treatment, A–D ) or treatment started when tumours are palpable (7–10 days) named “curative treatment” ( E, F ). Evolution of tumour volumes (mm 3 ) (A, C, E); follow-up of tumour progressions (B, D, F). * P

    Journal: PLoS ONE

    Article Title: Imatinib Mesylate Exerts Anti-Proliferative Effects on Osteosarcoma Cells and Inhibits the Tumour Growth in Immunocompetent Murine Models

    doi: 10.1371/journal.pone.0090795

    Figure Lengend Snippet: Imatinib mesylate inhibits osteosarcoma development in “preventive” and “curative” therapeutic context. Mice bearing undifferentiated POS-1 ( A, B ) or mixed osteoblastic/osteolytic MOS-J ( C–F ) osteosarcoma tumours (n = 8 per group) were assigned as control (vehicle), or imatinib mesylate (25, 50 or 100 mg/kg, daily oral administration). The treatment started 1 day after tumour cell inoculation (« preventive » treatment, A–D ) or treatment started when tumours are palpable (7–10 days) named “curative treatment” ( E, F ). Evolution of tumour volumes (mm 3 ) (A, C, E); follow-up of tumour progressions (B, D, F). * P

    Article Snippet: Imatinib mesylate (Gleevec, Novartis Pharma), a tyrosine kinase inhibitor, was originally developed for the treatment of chronic myeloid leukemia.

    Techniques: Mouse Assay

    Imatinib mesylate affects osteosarcoma cell proliferation by inducing a cell cycle arrest. Cell cycle distribution of human, mouse and rat osteosarcoma cell lines treated or not with imatinib mesylated for 48 hours was analyzed by propidium iodide staining and flow cytometry. All experiments were repeated 3 times, and representative results are shown.

    Journal: PLoS ONE

    Article Title: Imatinib Mesylate Exerts Anti-Proliferative Effects on Osteosarcoma Cells and Inhibits the Tumour Growth in Immunocompetent Murine Models

    doi: 10.1371/journal.pone.0090795

    Figure Lengend Snippet: Imatinib mesylate affects osteosarcoma cell proliferation by inducing a cell cycle arrest. Cell cycle distribution of human, mouse and rat osteosarcoma cell lines treated or not with imatinib mesylated for 48 hours was analyzed by propidium iodide staining and flow cytometry. All experiments were repeated 3 times, and representative results are shown.

    Article Snippet: Imatinib mesylate (Gleevec, Novartis Pharma), a tyrosine kinase inhibitor, was originally developed for the treatment of chronic myeloid leukemia.

    Techniques: Staining, Flow Cytometry, Cytometry

    Inhibitory effect of imatinib mesylate on osteosarcoma cell mitosis. Human MG63 ( A ), mouse MOS-J ( B ) and OSRGA ( C ) osteosarcoma cells were cultured in the presence or absence of imatinib mesylate with 25 µM, 20 µM and 10 µM respectively. Phase-contrast photos were taken every 10 minutes for 72 hours and the number of cell mitosis manually scored in a time-dependent manner.

    Journal: PLoS ONE

    Article Title: Imatinib Mesylate Exerts Anti-Proliferative Effects on Osteosarcoma Cells and Inhibits the Tumour Growth in Immunocompetent Murine Models

    doi: 10.1371/journal.pone.0090795

    Figure Lengend Snippet: Inhibitory effect of imatinib mesylate on osteosarcoma cell mitosis. Human MG63 ( A ), mouse MOS-J ( B ) and OSRGA ( C ) osteosarcoma cells were cultured in the presence or absence of imatinib mesylate with 25 µM, 20 µM and 10 µM respectively. Phase-contrast photos were taken every 10 minutes for 72 hours and the number of cell mitosis manually scored in a time-dependent manner.

    Article Snippet: Imatinib mesylate (Gleevec, Novartis Pharma), a tyrosine kinase inhibitor, was originally developed for the treatment of chronic myeloid leukemia.

    Techniques: Cell Culture

    Imatinib mesylate inhibits the PDGF-BB induced signalling pathways. Human, mouse and rat osteosarcoma were treated with 50/mL of PDGF-BB for 5 minutes in the presence of the absence of 25 µM of imatinib mesylate. PDGFRα, PDGFRβ, Akt, ERK1/2 phoshorylations were analyzed by Western blot compared to the levels of total forms of proteins and the levels of actin.

    Journal: PLoS ONE

    Article Title: Imatinib Mesylate Exerts Anti-Proliferative Effects on Osteosarcoma Cells and Inhibits the Tumour Growth in Immunocompetent Murine Models

    doi: 10.1371/journal.pone.0090795

    Figure Lengend Snippet: Imatinib mesylate inhibits the PDGF-BB induced signalling pathways. Human, mouse and rat osteosarcoma were treated with 50/mL of PDGF-BB for 5 minutes in the presence of the absence of 25 µM of imatinib mesylate. PDGFRα, PDGFRβ, Akt, ERK1/2 phoshorylations were analyzed by Western blot compared to the levels of total forms of proteins and the levels of actin.

    Article Snippet: Imatinib mesylate (Gleevec, Novartis Pharma), a tyrosine kinase inhibitor, was originally developed for the treatment of chronic myeloid leukemia.

    Techniques: Western Blot

    Imatinib mesylate inhibits AKT/mTOR signaling pathway in osteosarcoma cells and activates ERK1/2 phosphorylation: PDGFRα, a key target of osteosarcoma cells. Human, mouse and rat osteosarcoma were treated with various doses of imatinib mesylate to analyse the effects of the drug on intra-cellular signaling pathways. ( A ) Imatinib mesylate inhibits mTOR and Akt phosphorylation in human HOS and mouse MOS-J cells. RAD001 named everolimus (RAD), a mTOR inhibitor was used as a positive control. ( B ) Human Phospho-receptor tyrosine kinase array Kit was used to identify the molecular targets of imatinib mesylate. (C) Expression of PDGFRα and PDGFRβ analyzed in human, mouse and rat osteosarcoma cells by semi-quantitative RT-PCR.

    Journal: PLoS ONE

    Article Title: Imatinib Mesylate Exerts Anti-Proliferative Effects on Osteosarcoma Cells and Inhibits the Tumour Growth in Immunocompetent Murine Models

    doi: 10.1371/journal.pone.0090795

    Figure Lengend Snippet: Imatinib mesylate inhibits AKT/mTOR signaling pathway in osteosarcoma cells and activates ERK1/2 phosphorylation: PDGFRα, a key target of osteosarcoma cells. Human, mouse and rat osteosarcoma were treated with various doses of imatinib mesylate to analyse the effects of the drug on intra-cellular signaling pathways. ( A ) Imatinib mesylate inhibits mTOR and Akt phosphorylation in human HOS and mouse MOS-J cells. RAD001 named everolimus (RAD), a mTOR inhibitor was used as a positive control. ( B ) Human Phospho-receptor tyrosine kinase array Kit was used to identify the molecular targets of imatinib mesylate. (C) Expression of PDGFRα and PDGFRβ analyzed in human, mouse and rat osteosarcoma cells by semi-quantitative RT-PCR.

    Article Snippet: Imatinib mesylate (Gleevec, Novartis Pharma), a tyrosine kinase inhibitor, was originally developed for the treatment of chronic myeloid leukemia.

    Techniques: Positive Control, Expressing, Quantitative RT-PCR

    Osteosarcoma cell death induced by imatinib mesylate is partly dependent of caspase activity. ( A ) Human (MG63, HOS), mouse (POS-1, MOS-J) and rat osteosarcoma cells were cultured with or without increasing concentrations of imatinib mesylate. After 48 days of treatment, the alive and dead cells number was manually scored (from trypsinized and floating cells) after trypan blue exclusion. ( B ) Using similar culture conditions, caspase-3 activity was assessed using a kit CaspACE Assay System (Promega, USA). ( C ) On the same way, the involvement of caspase activity in cell death induced by imatinib mesylate was analyzed using the 2,3-bis(2 methoxy-4 nitro-5-sulfophenyl)2H-tetrazolium-5-carboxanilide (XTT) assay in the presence or the absence of 50 µM pan-caspase inhibitor Z-Vad-FMK(ZVAD).*p

    Journal: PLoS ONE

    Article Title: Imatinib Mesylate Exerts Anti-Proliferative Effects on Osteosarcoma Cells and Inhibits the Tumour Growth in Immunocompetent Murine Models

    doi: 10.1371/journal.pone.0090795

    Figure Lengend Snippet: Osteosarcoma cell death induced by imatinib mesylate is partly dependent of caspase activity. ( A ) Human (MG63, HOS), mouse (POS-1, MOS-J) and rat osteosarcoma cells were cultured with or without increasing concentrations of imatinib mesylate. After 48 days of treatment, the alive and dead cells number was manually scored (from trypsinized and floating cells) after trypan blue exclusion. ( B ) Using similar culture conditions, caspase-3 activity was assessed using a kit CaspACE Assay System (Promega, USA). ( C ) On the same way, the involvement of caspase activity in cell death induced by imatinib mesylate was analyzed using the 2,3-bis(2 methoxy-4 nitro-5-sulfophenyl)2H-tetrazolium-5-carboxanilide (XTT) assay in the presence or the absence of 50 µM pan-caspase inhibitor Z-Vad-FMK(ZVAD).*p

    Article Snippet: Imatinib mesylate (Gleevec, Novartis Pharma), a tyrosine kinase inhibitor, was originally developed for the treatment of chronic myeloid leukemia.

    Techniques: Activity Assay, Cell Culture, XTT Assay

    Imatinib mesylate increases osteosarcoma cell death. A kinetic of human (MG63), mouse (MOS-J) and rat (OSRGA) osteosarcoma cell death was analyzed by time-lapse microscopy in the presence or the absence of 25 µM, 20 µM or 10 µM imatinib mesylate respectively. The number of cell death was manually scored every 10 minutes until 72 hours.

    Journal: PLoS ONE

    Article Title: Imatinib Mesylate Exerts Anti-Proliferative Effects on Osteosarcoma Cells and Inhibits the Tumour Growth in Immunocompetent Murine Models

    doi: 10.1371/journal.pone.0090795

    Figure Lengend Snippet: Imatinib mesylate increases osteosarcoma cell death. A kinetic of human (MG63), mouse (MOS-J) and rat (OSRGA) osteosarcoma cell death was analyzed by time-lapse microscopy in the presence or the absence of 25 µM, 20 µM or 10 µM imatinib mesylate respectively. The number of cell death was manually scored every 10 minutes until 72 hours.

    Article Snippet: Imatinib mesylate (Gleevec, Novartis Pharma), a tyrosine kinase inhibitor, was originally developed for the treatment of chronic myeloid leukemia.

    Techniques: Time-lapse Microscopy

    Deletion of PHD2 in osteocytes stimulates angiogenesis. a , b CD31 immunostaining ( a ) of the tibial metaphysis with quantification ( b ) of blood vessel number and size in 8-week-old mice ( n = 8 Phd2 ot+ –10 Phd2 ot − ). c , d CD31 immunostaining ( c ) of the cortical diaphysis of tibiae with quantification ( d ) of blood vessel number and size ( n = 8 Phd2 ot+ –10 Phd2 ot− ). e Vegf and Plgf mRNA levels in femora of 8-week-old mice ( n = 8 Phd2 ot+ –10 Phd2 ot− ) and in vehicle (IDG VEH ) or IOX2-treated IDG-SW3 (IDG IOX2 ) cells ( n = 3). f Scheme of experimental design. HIF-1α (shH1) or HIF-2α (shH2) were silenced in IDG IOX2 cells and a scrambled shRNA (shScr) was used as control. Conditioned medium was subsequently used to culture HUVECs in monoculture or in co-culture with IDG-SW3 cells. g Proliferation of HUVECs in monoculture according to the scheme in f ( n = 3). h , i Human CD31 (hCD31) immunostaining ( h ) and quantification ( i ) of hCD31-positive surface in HUVECs co-cultured with IDG-SW3 cells, according to the scheme in f . When indicated, cells were treated with anti-VEGF 164 antibody or recombinant mouse soluble VEGFR-1 (sFlt) ( n = 3). Data are means ± SEM. * p

    Journal: Nature Communications

    Article Title: Osteocytic oxygen sensing controls bone mass through epigenetic regulation of sclerostin

    doi: 10.1038/s41467-018-04679-7

    Figure Lengend Snippet: Deletion of PHD2 in osteocytes stimulates angiogenesis. a , b CD31 immunostaining ( a ) of the tibial metaphysis with quantification ( b ) of blood vessel number and size in 8-week-old mice ( n = 8 Phd2 ot+ –10 Phd2 ot − ). c , d CD31 immunostaining ( c ) of the cortical diaphysis of tibiae with quantification ( d ) of blood vessel number and size ( n = 8 Phd2 ot+ –10 Phd2 ot− ). e Vegf and Plgf mRNA levels in femora of 8-week-old mice ( n = 8 Phd2 ot+ –10 Phd2 ot− ) and in vehicle (IDG VEH ) or IOX2-treated IDG-SW3 (IDG IOX2 ) cells ( n = 3). f Scheme of experimental design. HIF-1α (shH1) or HIF-2α (shH2) were silenced in IDG IOX2 cells and a scrambled shRNA (shScr) was used as control. Conditioned medium was subsequently used to culture HUVECs in monoculture or in co-culture with IDG-SW3 cells. g Proliferation of HUVECs in monoculture according to the scheme in f ( n = 3). h , i Human CD31 (hCD31) immunostaining ( h ) and quantification ( i ) of hCD31-positive surface in HUVECs co-cultured with IDG-SW3 cells, according to the scheme in f . When indicated, cells were treated with anti-VEGF 164 antibody or recombinant mouse soluble VEGFR-1 (sFlt) ( n = 3). Data are means ± SEM. * p

    Article Snippet: After culture, the endothelial cells were visualized by staining with a mouse-anti-human CD31 primary antibody (1/1000; Dako) and the TSA Cyanine 3 System (PerkinElmer).

    Techniques: Immunostaining, Mouse Assay, shRNA, Co-Culture Assay, Cell Culture, Recombinant

    FGF23 stimulation by hypoxia in vitro and in vivo. ( A ) FGF23 mRNA was significantly elevated in UMR-106 cells cultured under hypoxic conditions for 24 to 48 hours. In parallel, immunoblots (chart inset) for HIF1α (upper panel) showed an increase

    Journal: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

    Article Title: Neonatal Iron Deficiency Causes Abnormal Phosphate Metabolism by Elevating FGF23 in Normal and ADHR Mice

    doi: 10.1002/jbmr.2049

    Figure Lengend Snippet: FGF23 stimulation by hypoxia in vitro and in vivo. ( A ) FGF23 mRNA was significantly elevated in UMR-106 cells cultured under hypoxic conditions for 24 to 48 hours. In parallel, immunoblots (chart inset) for HIF1α (upper panel) showed an increase

    Article Snippet: We have also demonstrated that HIF activity could also be induced in parallel with FGF23 using the iron chelator DFO. Previous studies using UMR-106 cells demonstrated that other osteoblast/osteocyte genes can be controlled by hypoxia, including SOST, which is significantly reduced in parallel with increased expression and nuclear localization of activated β-catenin. In agreement, studies support that DMP1, MEPE, Connexin-43, and FGF23 may be controlled by hypoxia in MC3T3 cells and primary cultures of osteoblasts. Potentially, the interconnected control of these genes functions as a portion of a tissue and cell maturation program that may be required to balance mineralization through systemic phosphate control (FGF23), local osteoblast function (SOST, Connexin-43), and matrix deposition (DMP1, MEPE).

    Techniques: In Vitro, In Vivo, Cell Culture, Western Blot