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  • 99
    Thermo Fisher oligofectamine
    Oligofectamine, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 18955 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/oligofectamine/product/Thermo Fisher
    Average 99 stars, based on 18955 article reviews
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    oligofectamine - by Bioz Stars, 2020-08
    99/100 stars
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    99
    Millipore sirna resistant d2r
    <t>D2R</t> regulates spine development in hippocampal neurons in vivo . Hippocampal slices were prepared for diolistic labeling from 3–week–old C57BL/6 mice intraperitoneally injected with vehicle or with agonists or antagonists of D2R and D1R (a, c), or 4–week–old C57BL/6 mice injected with lentivirus expressing EGFP , D1R , D2R , D1R <t>siRNA</t> or D2R siRNA (b, d, e, f). (a, b) Representative images of DiI–labeled (a) or virus transduced (b) basal dendrites of hippocampal CA1 neurons. (c) Quantification of spine density for (a). (d) Quantification of spine density for (b). n = 3 mice for each condition, and 15 neurons from 3 slices of each animal were imaged for spine analysis in (c) and (d). (e) Sample trace of mEPSCs. (f) Analysis of mEPSC frequency; 5–8 slices from 3–5 animals were used for each condition; n = the total number of recorded neurons. Quinpirole: 0.5 mg/kg; bromocriptine: 10 mg/kg; eticlopride: 0.5 mg/kg; SKF38393: 1 mg/kg; SCH23390: 1 mg/kg. Data are presented as mean ± SEM. Scale bars, 5 μm. Two–tailed Mann–Whitney test was used to calculate p–values.
    Sirna Resistant D2r, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    TaKaRa piresneo
    <t>D2R</t> regulates spine development in hippocampal neurons in vivo . Hippocampal slices were prepared for diolistic labeling from 3–week–old C57BL/6 mice intraperitoneally injected with vehicle or with agonists or antagonists of D2R and D1R (a, c), or 4–week–old C57BL/6 mice injected with lentivirus expressing EGFP , D1R , D2R , D1R <t>siRNA</t> or D2R siRNA (b, d, e, f). (a, b) Representative images of DiI–labeled (a) or virus transduced (b) basal dendrites of hippocampal CA1 neurons. (c) Quantification of spine density for (a). (d) Quantification of spine density for (b). n = 3 mice for each condition, and 15 neurons from 3 slices of each animal were imaged for spine analysis in (c) and (d). (e) Sample trace of mEPSCs. (f) Analysis of mEPSC frequency; 5–8 slices from 3–5 animals were used for each condition; n = the total number of recorded neurons. Quinpirole: 0.5 mg/kg; bromocriptine: 10 mg/kg; eticlopride: 0.5 mg/kg; SKF38393: 1 mg/kg; SCH23390: 1 mg/kg. Data are presented as mean ± SEM. Scale bars, 5 μm. Two–tailed Mann–Whitney test was used to calculate p–values.
    Piresneo, supplied by TaKaRa, used in various techniques. Bioz Stars score: 89/100, based on 319 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 89 stars, based on 319 article reviews
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    piresneo - by Bioz Stars, 2020-08
    89/100 stars
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    89
    Thermo Fisher pcdm8
    <t>D2R</t> regulates spine development in hippocampal neurons in vivo . Hippocampal slices were prepared for diolistic labeling from 3–week–old C57BL/6 mice intraperitoneally injected with vehicle or with agonists or antagonists of D2R and D1R (a, c), or 4–week–old C57BL/6 mice injected with lentivirus expressing EGFP , D1R , D2R , D1R <t>siRNA</t> or D2R siRNA (b, d, e, f). (a, b) Representative images of DiI–labeled (a) or virus transduced (b) basal dendrites of hippocampal CA1 neurons. (c) Quantification of spine density for (a). (d) Quantification of spine density for (b). n = 3 mice for each condition, and 15 neurons from 3 slices of each animal were imaged for spine analysis in (c) and (d). (e) Sample trace of mEPSCs. (f) Analysis of mEPSC frequency; 5–8 slices from 3–5 animals were used for each condition; n = the total number of recorded neurons. Quinpirole: 0.5 mg/kg; bromocriptine: 10 mg/kg; eticlopride: 0.5 mg/kg; SKF38393: 1 mg/kg; SCH23390: 1 mg/kg. Data are presented as mean ± SEM. Scale bars, 5 μm. Two–tailed Mann–Whitney test was used to calculate p–values.
    Pcdm8, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 176 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Santa Cruz Biotechnology fatp4
    Angpt2–ITGα5β1 signaling facilitates FA transport through CD36 and FATP3. a , c , d , g – p HUVECs were treated with vehicle or Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) for 15 min or indicated time points. a Comparisons of short-chain FA (BODIPY C-5, 8 μM; n = 4) and long-chain FA (BODIPY C-12 and C-16, 8 μM; n = 4 for vehicle and 3 for Mn 2+ + Angpt2) uptake. b Comparisons of depletion efficiency of CD36 , FATP3 , and <t>FATP4</t> in corresponding siRNA treated HUVECs. n = 3. c , d Comparisons and representative images of FA (BODIPY C-12, 8 μM) uptake in siControl ( n = 7, 5), si CD36 ( n = 4, 5), si FATP3 ( n = 6), or si FATP4 ( n = 6, 8) HUVECs. Scale bars, 30 μm. e Diagram depicting the endothelial FA transport assay. HUVECs were cultured until confluence on trans-wells and FA (BODIPY C-12, 8 μM) was added to upper layer of trans-well, followed by analysis of transported FA in bottom well. f Comparisons of FA transport for indicated time points after Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) treatment in siControl ( n = 3 for 10, 30 min and 4 for 20 min), si FATP3 ( n = 3) or si FATP4 ( n = 4) HUVECs. n = 3–5. g Comparison of mRNA expression of CD36 and FATP3 . h Representative images of FATP3-td-Tomato transfected HUVECs. Scale bars, 50 μm. i Representative images of CD36 and ITGβ1 expression. Expression of active ITGβ1 (arrows) and CD36 (arrowheads) are co-localized after Angpt2 treatment. Scale bars, 50 μm. j , k Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex. Scale bars, 20 μm. l – n Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex in HUVECs pre-treated with vehicle or ITGα5β1 blocking peptide (ATN-161, 10 μM) for 15 min. Scale bars, 50 μm ( l ); 20 μm ( m ). o , p Immunoprecipitation with anti-IgG and anti-CD36 antibody in HUVECs. Immunoblot analysis with anti-ITGβ1 and anti-ITGα5 antibodies are shown. Graph indicates normalized ratio of immunoprecipitated ITGβ1 per input. Unless otherwise denoted, each dot indicates a mean of triplicate values from three independent experiments and horizontal bars indicate mean ± SD or SEM ( f ) and P values versus control by two-tailed Student’s t test. NS not significant.
    Fatp4, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 90 stars, based on 34 article reviews
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    85
    Carl Zeiss confocal fluorescent microscope lsm 510 meta
    Angpt2–ITGα5β1 signaling facilitates FA transport through CD36 and FATP3. a , c , d , g – p HUVECs were treated with vehicle or Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) for 15 min or indicated time points. a Comparisons of short-chain FA (BODIPY C-5, 8 μM; n = 4) and long-chain FA (BODIPY C-12 and C-16, 8 μM; n = 4 for vehicle and 3 for Mn 2+ + Angpt2) uptake. b Comparisons of depletion efficiency of CD36 , FATP3 , and <t>FATP4</t> in corresponding siRNA treated HUVECs. n = 3. c , d Comparisons and representative images of FA (BODIPY C-12, 8 μM) uptake in siControl ( n = 7, 5), si CD36 ( n = 4, 5), si FATP3 ( n = 6), or si FATP4 ( n = 6, 8) HUVECs. Scale bars, 30 μm. e Diagram depicting the endothelial FA transport assay. HUVECs were cultured until confluence on trans-wells and FA (BODIPY C-12, 8 μM) was added to upper layer of trans-well, followed by analysis of transported FA in bottom well. f Comparisons of FA transport for indicated time points after Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) treatment in siControl ( n = 3 for 10, 30 min and 4 for 20 min), si FATP3 ( n = 3) or si FATP4 ( n = 4) HUVECs. n = 3–5. g Comparison of mRNA expression of CD36 and FATP3 . h Representative images of FATP3-td-Tomato transfected HUVECs. Scale bars, 50 μm. i Representative images of CD36 and ITGβ1 expression. Expression of active ITGβ1 (arrows) and CD36 (arrowheads) are co-localized after Angpt2 treatment. Scale bars, 50 μm. j , k Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex. Scale bars, 20 μm. l – n Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex in HUVECs pre-treated with vehicle or ITGα5β1 blocking peptide (ATN-161, 10 μM) for 15 min. Scale bars, 50 μm ( l ); 20 μm ( m ). o , p Immunoprecipitation with anti-IgG and anti-CD36 antibody in HUVECs. Immunoblot analysis with anti-ITGβ1 and anti-ITGα5 antibodies are shown. Graph indicates normalized ratio of immunoprecipitated ITGβ1 per input. Unless otherwise denoted, each dot indicates a mean of triplicate values from three independent experiments and horizontal bars indicate mean ± SD or SEM ( f ) and P values versus control by two-tailed Student’s t test. NS not significant.
    Confocal Fluorescent Microscope Lsm 510 Meta, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 85/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Becton Dickinson facscan flow cytometer
    Angpt2–ITGα5β1 signaling facilitates FA transport through CD36 and FATP3. a , c , d , g – p HUVECs were treated with vehicle or Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) for 15 min or indicated time points. a Comparisons of short-chain FA (BODIPY C-5, 8 μM; n = 4) and long-chain FA (BODIPY C-12 and C-16, 8 μM; n = 4 for vehicle and 3 for Mn 2+ + Angpt2) uptake. b Comparisons of depletion efficiency of CD36 , FATP3 , and <t>FATP4</t> in corresponding siRNA treated HUVECs. n = 3. c , d Comparisons and representative images of FA (BODIPY C-12, 8 μM) uptake in siControl ( n = 7, 5), si CD36 ( n = 4, 5), si FATP3 ( n = 6), or si FATP4 ( n = 6, 8) HUVECs. Scale bars, 30 μm. e Diagram depicting the endothelial FA transport assay. HUVECs were cultured until confluence on trans-wells and FA (BODIPY C-12, 8 μM) was added to upper layer of trans-well, followed by analysis of transported FA in bottom well. f Comparisons of FA transport for indicated time points after Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) treatment in siControl ( n = 3 for 10, 30 min and 4 for 20 min), si FATP3 ( n = 3) or si FATP4 ( n = 4) HUVECs. n = 3–5. g Comparison of mRNA expression of CD36 and FATP3 . h Representative images of FATP3-td-Tomato transfected HUVECs. Scale bars, 50 μm. i Representative images of CD36 and ITGβ1 expression. Expression of active ITGβ1 (arrows) and CD36 (arrowheads) are co-localized after Angpt2 treatment. Scale bars, 50 μm. j , k Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex. Scale bars, 20 μm. l – n Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex in HUVECs pre-treated with vehicle or ITGα5β1 blocking peptide (ATN-161, 10 μM) for 15 min. Scale bars, 50 μm ( l ); 20 μm ( m ). o , p Immunoprecipitation with anti-IgG and anti-CD36 antibody in HUVECs. Immunoblot analysis with anti-ITGβ1 and anti-ITGα5 antibodies are shown. Graph indicates normalized ratio of immunoprecipitated ITGβ1 per input. Unless otherwise denoted, each dot indicates a mean of triplicate values from three independent experiments and horizontal bars indicate mean ± SD or SEM ( f ) and P values versus control by two-tailed Student’s t test. NS not significant.
    Facscan Flow Cytometer, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 94/100, based on 46948 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    facscan flow cytometer - by Bioz Stars, 2020-08
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    93
    Molecular Devices LLC microplate reader
    Angpt2–ITGα5β1 signaling facilitates FA transport through CD36 and FATP3. a , c , d , g – p HUVECs were treated with vehicle or Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) for 15 min or indicated time points. a Comparisons of short-chain FA (BODIPY C-5, 8 μM; n = 4) and long-chain FA (BODIPY C-12 and C-16, 8 μM; n = 4 for vehicle and 3 for Mn 2+ + Angpt2) uptake. b Comparisons of depletion efficiency of CD36 , FATP3 , and <t>FATP4</t> in corresponding siRNA treated HUVECs. n = 3. c , d Comparisons and representative images of FA (BODIPY C-12, 8 μM) uptake in siControl ( n = 7, 5), si CD36 ( n = 4, 5), si FATP3 ( n = 6), or si FATP4 ( n = 6, 8) HUVECs. Scale bars, 30 μm. e Diagram depicting the endothelial FA transport assay. HUVECs were cultured until confluence on trans-wells and FA (BODIPY C-12, 8 μM) was added to upper layer of trans-well, followed by analysis of transported FA in bottom well. f Comparisons of FA transport for indicated time points after Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) treatment in siControl ( n = 3 for 10, 30 min and 4 for 20 min), si FATP3 ( n = 3) or si FATP4 ( n = 4) HUVECs. n = 3–5. g Comparison of mRNA expression of CD36 and FATP3 . h Representative images of FATP3-td-Tomato transfected HUVECs. Scale bars, 50 μm. i Representative images of CD36 and ITGβ1 expression. Expression of active ITGβ1 (arrows) and CD36 (arrowheads) are co-localized after Angpt2 treatment. Scale bars, 50 μm. j , k Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex. Scale bars, 20 μm. l – n Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex in HUVECs pre-treated with vehicle or ITGα5β1 blocking peptide (ATN-161, 10 μM) for 15 min. Scale bars, 50 μm ( l ); 20 μm ( m ). o , p Immunoprecipitation with anti-IgG and anti-CD36 antibody in HUVECs. Immunoblot analysis with anti-ITGβ1 and anti-ITGα5 antibodies are shown. Graph indicates normalized ratio of immunoprecipitated ITGβ1 per input. Unless otherwise denoted, each dot indicates a mean of triplicate values from three independent experiments and horizontal bars indicate mean ± SD or SEM ( f ) and P values versus control by two-tailed Student’s t test. NS not significant.
    Microplate Reader, supplied by Molecular Devices LLC, used in various techniques. Bioz Stars score: 93/100, based on 28124 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/microplate reader/product/Molecular Devices LLC
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    microplate reader - by Bioz Stars, 2020-08
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    91
    Bioneer Corporation itgβ3
    Angpt2–ITGα5β1 signaling facilitates FA transport through CD36 and FATP3. a , c , d , g – p HUVECs were treated with vehicle or Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) for 15 min or indicated time points. a Comparisons of short-chain FA (BODIPY C-5, 8 μM; n = 4) and long-chain FA (BODIPY C-12 and C-16, 8 μM; n = 4 for vehicle and 3 for Mn 2+ + Angpt2) uptake. b Comparisons of depletion efficiency of CD36 , FATP3 , and <t>FATP4</t> in corresponding siRNA treated HUVECs. n = 3. c , d Comparisons and representative images of FA (BODIPY C-12, 8 μM) uptake in siControl ( n = 7, 5), si CD36 ( n = 4, 5), si FATP3 ( n = 6), or si FATP4 ( n = 6, 8) HUVECs. Scale bars, 30 μm. e Diagram depicting the endothelial FA transport assay. HUVECs were cultured until confluence on trans-wells and FA (BODIPY C-12, 8 μM) was added to upper layer of trans-well, followed by analysis of transported FA in bottom well. f Comparisons of FA transport for indicated time points after Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) treatment in siControl ( n = 3 for 10, 30 min and 4 for 20 min), si FATP3 ( n = 3) or si FATP4 ( n = 4) HUVECs. n = 3–5. g Comparison of mRNA expression of CD36 and FATP3 . h Representative images of FATP3-td-Tomato transfected HUVECs. Scale bars, 50 μm. i Representative images of CD36 and ITGβ1 expression. Expression of active ITGβ1 (arrows) and CD36 (arrowheads) are co-localized after Angpt2 treatment. Scale bars, 50 μm. j , k Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex. Scale bars, 20 μm. l – n Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex in HUVECs pre-treated with vehicle or ITGα5β1 blocking peptide (ATN-161, 10 μM) for 15 min. Scale bars, 50 μm ( l ); 20 μm ( m ). o , p Immunoprecipitation with anti-IgG and anti-CD36 antibody in HUVECs. Immunoblot analysis with anti-ITGβ1 and anti-ITGα5 antibodies are shown. Graph indicates normalized ratio of immunoprecipitated ITGβ1 per input. Unless otherwise denoted, each dot indicates a mean of triplicate values from three independent experiments and horizontal bars indicate mean ± SD or SEM ( f ) and P values versus control by two-tailed Student’s t test. NS not significant.
    Itgβ3, supplied by Bioneer Corporation, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/itgβ3/product/Bioneer Corporation
    Average 91 stars, based on 1 article reviews
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    itgβ3 - by Bioz Stars, 2020-08
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    91
    PerkinElmer victor3 1420 multilabel counter
    Angpt2–ITGα5β1 signaling facilitates FA transport through CD36 and FATP3. a , c , d , g – p HUVECs were treated with vehicle or Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) for 15 min or indicated time points. a Comparisons of short-chain FA (BODIPY C-5, 8 μM; n = 4) and long-chain FA (BODIPY C-12 and C-16, 8 μM; n = 4 for vehicle and 3 for Mn 2+ + Angpt2) uptake. b Comparisons of depletion efficiency of CD36 , FATP3 , and <t>FATP4</t> in corresponding siRNA treated HUVECs. n = 3. c , d Comparisons and representative images of FA (BODIPY C-12, 8 μM) uptake in siControl ( n = 7, 5), si CD36 ( n = 4, 5), si FATP3 ( n = 6), or si FATP4 ( n = 6, 8) HUVECs. Scale bars, 30 μm. e Diagram depicting the endothelial FA transport assay. HUVECs were cultured until confluence on trans-wells and FA (BODIPY C-12, 8 μM) was added to upper layer of trans-well, followed by analysis of transported FA in bottom well. f Comparisons of FA transport for indicated time points after Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) treatment in siControl ( n = 3 for 10, 30 min and 4 for 20 min), si FATP3 ( n = 3) or si FATP4 ( n = 4) HUVECs. n = 3–5. g Comparison of mRNA expression of CD36 and FATP3 . h Representative images of FATP3-td-Tomato transfected HUVECs. Scale bars, 50 μm. i Representative images of CD36 and ITGβ1 expression. Expression of active ITGβ1 (arrows) and CD36 (arrowheads) are co-localized after Angpt2 treatment. Scale bars, 50 μm. j , k Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex. Scale bars, 20 μm. l – n Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex in HUVECs pre-treated with vehicle or ITGα5β1 blocking peptide (ATN-161, 10 μM) for 15 min. Scale bars, 50 μm ( l ); 20 μm ( m ). o , p Immunoprecipitation with anti-IgG and anti-CD36 antibody in HUVECs. Immunoblot analysis with anti-ITGβ1 and anti-ITGα5 antibodies are shown. Graph indicates normalized ratio of immunoprecipitated ITGβ1 per input. Unless otherwise denoted, each dot indicates a mean of triplicate values from three independent experiments and horizontal bars indicate mean ± SD or SEM ( f ) and P values versus control by two-tailed Student’s t test. NS not significant.
    Victor3 1420 Multilabel Counter, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 91/100, based on 743 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/victor3 1420 multilabel counter/product/PerkinElmer
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    99
    Thermo Fisher fluoroskan ascent fluorometer
    Angpt2–ITGα5β1 signaling facilitates FA transport through CD36 and FATP3. a , c , d , g – p HUVECs were treated with vehicle or Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) for 15 min or indicated time points. a Comparisons of short-chain FA (BODIPY C-5, 8 μM; n = 4) and long-chain FA (BODIPY C-12 and C-16, 8 μM; n = 4 for vehicle and 3 for Mn 2+ + Angpt2) uptake. b Comparisons of depletion efficiency of CD36 , FATP3 , and <t>FATP4</t> in corresponding siRNA treated HUVECs. n = 3. c , d Comparisons and representative images of FA (BODIPY C-12, 8 μM) uptake in siControl ( n = 7, 5), si CD36 ( n = 4, 5), si FATP3 ( n = 6), or si FATP4 ( n = 6, 8) HUVECs. Scale bars, 30 μm. e Diagram depicting the endothelial FA transport assay. HUVECs were cultured until confluence on trans-wells and FA (BODIPY C-12, 8 μM) was added to upper layer of trans-well, followed by analysis of transported FA in bottom well. f Comparisons of FA transport for indicated time points after Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) treatment in siControl ( n = 3 for 10, 30 min and 4 for 20 min), si FATP3 ( n = 3) or si FATP4 ( n = 4) HUVECs. n = 3–5. g Comparison of mRNA expression of CD36 and FATP3 . h Representative images of FATP3-td-Tomato transfected HUVECs. Scale bars, 50 μm. i Representative images of CD36 and ITGβ1 expression. Expression of active ITGβ1 (arrows) and CD36 (arrowheads) are co-localized after Angpt2 treatment. Scale bars, 50 μm. j , k Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex. Scale bars, 20 μm. l – n Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex in HUVECs pre-treated with vehicle or ITGα5β1 blocking peptide (ATN-161, 10 μM) for 15 min. Scale bars, 50 μm ( l ); 20 μm ( m ). o , p Immunoprecipitation with anti-IgG and anti-CD36 antibody in HUVECs. Immunoblot analysis with anti-ITGβ1 and anti-ITGα5 antibodies are shown. Graph indicates normalized ratio of immunoprecipitated ITGβ1 per input. Unless otherwise denoted, each dot indicates a mean of triplicate values from three independent experiments and horizontal bars indicate mean ± SD or SEM ( f ) and P values versus control by two-tailed Student’s t test. NS not significant.
    Fluoroskan Ascent Fluorometer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 416 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fluoroskan ascent fluorometer/product/Thermo Fisher
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    fluoroskan ascent fluorometer - by Bioz Stars, 2020-08
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    91
    Bioneer Corporation itgβ5
    Angpt2–ITGα5β1 signaling facilitates FA transport through CD36 and FATP3. a , c , d , g – p HUVECs were treated with vehicle or Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) for 15 min or indicated time points. a Comparisons of short-chain FA (BODIPY C-5, 8 μM; n = 4) and long-chain FA (BODIPY C-12 and C-16, 8 μM; n = 4 for vehicle and 3 for Mn 2+ + Angpt2) uptake. b Comparisons of depletion efficiency of CD36 , FATP3 , and <t>FATP4</t> in corresponding siRNA treated HUVECs. n = 3. c , d Comparisons and representative images of FA (BODIPY C-12, 8 μM) uptake in siControl ( n = 7, 5), si CD36 ( n = 4, 5), si FATP3 ( n = 6), or si FATP4 ( n = 6, 8) HUVECs. Scale bars, 30 μm. e Diagram depicting the endothelial FA transport assay. HUVECs were cultured until confluence on trans-wells and FA (BODIPY C-12, 8 μM) was added to upper layer of trans-well, followed by analysis of transported FA in bottom well. f Comparisons of FA transport for indicated time points after Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) treatment in siControl ( n = 3 for 10, 30 min and 4 for 20 min), si FATP3 ( n = 3) or si FATP4 ( n = 4) HUVECs. n = 3–5. g Comparison of mRNA expression of CD36 and FATP3 . h Representative images of FATP3-td-Tomato transfected HUVECs. Scale bars, 50 μm. i Representative images of CD36 and ITGβ1 expression. Expression of active ITGβ1 (arrows) and CD36 (arrowheads) are co-localized after Angpt2 treatment. Scale bars, 50 μm. j , k Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex. Scale bars, 20 μm. l – n Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex in HUVECs pre-treated with vehicle or ITGα5β1 blocking peptide (ATN-161, 10 μM) for 15 min. Scale bars, 50 μm ( l ); 20 μm ( m ). o , p Immunoprecipitation with anti-IgG and anti-CD36 antibody in HUVECs. Immunoblot analysis with anti-ITGβ1 and anti-ITGα5 antibodies are shown. Graph indicates normalized ratio of immunoprecipitated ITGβ1 per input. Unless otherwise denoted, each dot indicates a mean of triplicate values from three independent experiments and horizontal bars indicate mean ± SD or SEM ( f ) and P values versus control by two-tailed Student’s t test. NS not significant.
    Itgβ5, supplied by Bioneer Corporation, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/itgβ5/product/Bioneer Corporation
    Average 91 stars, based on 1 article reviews
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    itgβ5 - by Bioz Stars, 2020-08
    91/100 stars
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    Bioneer Corporation itgα5
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    Analysis of AAT Expression after Transfection with Liposomes or <t>Lipofectamine</t> 2000 Containing AAT-Encoding mRNA Concentration of AAT protein in cell supernatants after transfection with AAT mRNA encapsulated in liposomes or Lipofectamine 2000 for 24 or 72 hr. Data are shown as mean ± SEM (n = 5). **p
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    Image Search Results


    D2R regulates spine development in hippocampal neurons in vivo . Hippocampal slices were prepared for diolistic labeling from 3–week–old C57BL/6 mice intraperitoneally injected with vehicle or with agonists or antagonists of D2R and D1R (a, c), or 4–week–old C57BL/6 mice injected with lentivirus expressing EGFP , D1R , D2R , D1R siRNA or D2R siRNA (b, d, e, f). (a, b) Representative images of DiI–labeled (a) or virus transduced (b) basal dendrites of hippocampal CA1 neurons. (c) Quantification of spine density for (a). (d) Quantification of spine density for (b). n = 3 mice for each condition, and 15 neurons from 3 slices of each animal were imaged for spine analysis in (c) and (d). (e) Sample trace of mEPSCs. (f) Analysis of mEPSC frequency; 5–8 slices from 3–5 animals were used for each condition; n = the total number of recorded neurons. Quinpirole: 0.5 mg/kg; bromocriptine: 10 mg/kg; eticlopride: 0.5 mg/kg; SKF38393: 1 mg/kg; SCH23390: 1 mg/kg. Data are presented as mean ± SEM. Scale bars, 5 μm. Two–tailed Mann–Whitney test was used to calculate p–values.

    Journal: Nature neuroscience

    Article Title: Age-dependent regulation of synaptic connections by dopamine D2 receptors

    doi: 10.1038/nn.3542

    Figure Lengend Snippet: D2R regulates spine development in hippocampal neurons in vivo . Hippocampal slices were prepared for diolistic labeling from 3–week–old C57BL/6 mice intraperitoneally injected with vehicle or with agonists or antagonists of D2R and D1R (a, c), or 4–week–old C57BL/6 mice injected with lentivirus expressing EGFP , D1R , D2R , D1R siRNA or D2R siRNA (b, d, e, f). (a, b) Representative images of DiI–labeled (a) or virus transduced (b) basal dendrites of hippocampal CA1 neurons. (c) Quantification of spine density for (a). (d) Quantification of spine density for (b). n = 3 mice for each condition, and 15 neurons from 3 slices of each animal were imaged for spine analysis in (c) and (d). (e) Sample trace of mEPSCs. (f) Analysis of mEPSC frequency; 5–8 slices from 3–5 animals were used for each condition; n = the total number of recorded neurons. Quinpirole: 0.5 mg/kg; bromocriptine: 10 mg/kg; eticlopride: 0.5 mg/kg; SKF38393: 1 mg/kg; SCH23390: 1 mg/kg. Data are presented as mean ± SEM. Scale bars, 5 μm. Two–tailed Mann–Whitney test was used to calculate p–values.

    Article Snippet: Annealed oligos containing siRNA sequences targeted to rat D2R (D2R siRNA–1 : 1138′–CTCGGTGTGTTCATCATCT–1156′— a region conserved between rats and mice; D2R siRNA–2 : 1269′–CCCCATCATCTACACCACC–1; D2R siRNA–3 : 530′–CAGACCAGAATGAGTGTAT–548′) and mouse D1R (1075′–GAGACTGTAAGCATCAACA–1093′), and the cDNAs of myc –tagged D1R and HA –tagged D2R were inserted into the pSuper and the pRRLsin lentiviral vectors. cDNAs for D2R , Beclin1 , ATG5 and siRNA–resistant D2R (C1140G, G1146T, C1149T, C1155T, generated by mutagenesis using the KOD kit, Novagen) were cloned into the pGW1 vector behind the HA sequence.

    Techniques: In Vivo, Labeling, Mouse Assay, Injection, Expressing, Two Tailed Test, MANN-WHITNEY

    Spine deficiency in sandy mice is caused by D2R hyperactivity. (a–c) Primary hippocampal neurons were prepared from wild–type and sandy mice, transfected with the Venus construct at DIV14, and imaged at DIV17. Representative images of transfected neurons and dendrites at a higher magnification are shown in (a), and quantification is shown in (b, c); the results were replicated by three independent experiments, and one of the three replicates (n = 15 for each condition) is shown in the histograms. (d–i) At 3 weeks of age, sandy mice and their wild–type littermates were injected with lentivirus expressing EGFP or D2R siRNA, or intraperitoneally injected with vehicle or forskolin (4 mg/kg), then used for preparation of hippocampal slices for spine analysis (d, e), mEPSC analysis (f, g) or diolistic labeling (h, i). (d, h) Representative images of basal dendrites from CA1 neurons. (e) Quantification of spine density for (d); 15 neurons from 3 slices of each animal and 3 mice for each condition were imaged and analyzed. (i) Quantification of spine density for (h); 15 neurons from 3 slices of each animal and 3 mice for each condition were imaged and analyzed. (f) Sample trace of mEPSCs. (g) Quantificaiton for (f); 5–8 slices of 3–5 animals for each condition; n= the total number of recorded neurons. Scale bar, 20 μm for images of neurons and 5 μm for images of dendrites. Data are presented as mean ± SEM. Mann–Whitney test was used for statistical analysis.

    Journal: Nature neuroscience

    Article Title: Age-dependent regulation of synaptic connections by dopamine D2 receptors

    doi: 10.1038/nn.3542

    Figure Lengend Snippet: Spine deficiency in sandy mice is caused by D2R hyperactivity. (a–c) Primary hippocampal neurons were prepared from wild–type and sandy mice, transfected with the Venus construct at DIV14, and imaged at DIV17. Representative images of transfected neurons and dendrites at a higher magnification are shown in (a), and quantification is shown in (b, c); the results were replicated by three independent experiments, and one of the three replicates (n = 15 for each condition) is shown in the histograms. (d–i) At 3 weeks of age, sandy mice and their wild–type littermates were injected with lentivirus expressing EGFP or D2R siRNA, or intraperitoneally injected with vehicle or forskolin (4 mg/kg), then used for preparation of hippocampal slices for spine analysis (d, e), mEPSC analysis (f, g) or diolistic labeling (h, i). (d, h) Representative images of basal dendrites from CA1 neurons. (e) Quantification of spine density for (d); 15 neurons from 3 slices of each animal and 3 mice for each condition were imaged and analyzed. (i) Quantification of spine density for (h); 15 neurons from 3 slices of each animal and 3 mice for each condition were imaged and analyzed. (f) Sample trace of mEPSCs. (g) Quantificaiton for (f); 5–8 slices of 3–5 animals for each condition; n= the total number of recorded neurons. Scale bar, 20 μm for images of neurons and 5 μm for images of dendrites. Data are presented as mean ± SEM. Mann–Whitney test was used for statistical analysis.

    Article Snippet: Annealed oligos containing siRNA sequences targeted to rat D2R (D2R siRNA–1 : 1138′–CTCGGTGTGTTCATCATCT–1156′— a region conserved between rats and mice; D2R siRNA–2 : 1269′–CCCCATCATCTACACCACC–1; D2R siRNA–3 : 530′–CAGACCAGAATGAGTGTAT–548′) and mouse D1R (1075′–GAGACTGTAAGCATCAACA–1093′), and the cDNAs of myc –tagged D1R and HA –tagged D2R were inserted into the pSuper and the pRRLsin lentiviral vectors. cDNAs for D2R , Beclin1 , ATG5 and siRNA–resistant D2R (C1140G, G1146T, C1149T, C1155T, generated by mutagenesis using the KOD kit, Novagen) were cloned into the pGW1 vector behind the HA sequence.

    Techniques: Mouse Assay, Transfection, Construct, Injection, Expressing, Labeling, MANN-WHITNEY

    Angpt2–ITGα5β1 signaling facilitates FA transport through CD36 and FATP3. a , c , d , g – p HUVECs were treated with vehicle or Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) for 15 min or indicated time points. a Comparisons of short-chain FA (BODIPY C-5, 8 μM; n = 4) and long-chain FA (BODIPY C-12 and C-16, 8 μM; n = 4 for vehicle and 3 for Mn 2+ + Angpt2) uptake. b Comparisons of depletion efficiency of CD36 , FATP3 , and FATP4 in corresponding siRNA treated HUVECs. n = 3. c , d Comparisons and representative images of FA (BODIPY C-12, 8 μM) uptake in siControl ( n = 7, 5), si CD36 ( n = 4, 5), si FATP3 ( n = 6), or si FATP4 ( n = 6, 8) HUVECs. Scale bars, 30 μm. e Diagram depicting the endothelial FA transport assay. HUVECs were cultured until confluence on trans-wells and FA (BODIPY C-12, 8 μM) was added to upper layer of trans-well, followed by analysis of transported FA in bottom well. f Comparisons of FA transport for indicated time points after Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) treatment in siControl ( n = 3 for 10, 30 min and 4 for 20 min), si FATP3 ( n = 3) or si FATP4 ( n = 4) HUVECs. n = 3–5. g Comparison of mRNA expression of CD36 and FATP3 . h Representative images of FATP3-td-Tomato transfected HUVECs. Scale bars, 50 μm. i Representative images of CD36 and ITGβ1 expression. Expression of active ITGβ1 (arrows) and CD36 (arrowheads) are co-localized after Angpt2 treatment. Scale bars, 50 μm. j , k Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex. Scale bars, 20 μm. l – n Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex in HUVECs pre-treated with vehicle or ITGα5β1 blocking peptide (ATN-161, 10 μM) for 15 min. Scale bars, 50 μm ( l ); 20 μm ( m ). o , p Immunoprecipitation with anti-IgG and anti-CD36 antibody in HUVECs. Immunoblot analysis with anti-ITGβ1 and anti-ITGα5 antibodies are shown. Graph indicates normalized ratio of immunoprecipitated ITGβ1 per input. Unless otherwise denoted, each dot indicates a mean of triplicate values from three independent experiments and horizontal bars indicate mean ± SD or SEM ( f ) and P values versus control by two-tailed Student’s t test. NS not significant.

    Journal: Nature Communications

    Article Title: Angiopoietin-2–integrin α5β1 signaling enhances vascular fatty acid transport and prevents ectopic lipid-induced insulin resistance

    doi: 10.1038/s41467-020-16795-4

    Figure Lengend Snippet: Angpt2–ITGα5β1 signaling facilitates FA transport through CD36 and FATP3. a , c , d , g – p HUVECs were treated with vehicle or Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) for 15 min or indicated time points. a Comparisons of short-chain FA (BODIPY C-5, 8 μM; n = 4) and long-chain FA (BODIPY C-12 and C-16, 8 μM; n = 4 for vehicle and 3 for Mn 2+ + Angpt2) uptake. b Comparisons of depletion efficiency of CD36 , FATP3 , and FATP4 in corresponding siRNA treated HUVECs. n = 3. c , d Comparisons and representative images of FA (BODIPY C-12, 8 μM) uptake in siControl ( n = 7, 5), si CD36 ( n = 4, 5), si FATP3 ( n = 6), or si FATP4 ( n = 6, 8) HUVECs. Scale bars, 30 μm. e Diagram depicting the endothelial FA transport assay. HUVECs were cultured until confluence on trans-wells and FA (BODIPY C-12, 8 μM) was added to upper layer of trans-well, followed by analysis of transported FA in bottom well. f Comparisons of FA transport for indicated time points after Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) treatment in siControl ( n = 3 for 10, 30 min and 4 for 20 min), si FATP3 ( n = 3) or si FATP4 ( n = 4) HUVECs. n = 3–5. g Comparison of mRNA expression of CD36 and FATP3 . h Representative images of FATP3-td-Tomato transfected HUVECs. Scale bars, 50 μm. i Representative images of CD36 and ITGβ1 expression. Expression of active ITGβ1 (arrows) and CD36 (arrowheads) are co-localized after Angpt2 treatment. Scale bars, 50 μm. j , k Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex. Scale bars, 20 μm. l – n Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex in HUVECs pre-treated with vehicle or ITGα5β1 blocking peptide (ATN-161, 10 μM) for 15 min. Scale bars, 50 μm ( l ); 20 μm ( m ). o , p Immunoprecipitation with anti-IgG and anti-CD36 antibody in HUVECs. Immunoblot analysis with anti-ITGβ1 and anti-ITGα5 antibodies are shown. Graph indicates normalized ratio of immunoprecipitated ITGβ1 per input. Unless otherwise denoted, each dot indicates a mean of triplicate values from three independent experiments and horizontal bars indicate mean ± SD or SEM ( f ) and P values versus control by two-tailed Student’s t test. NS not significant.

    Article Snippet: siRNA transfectionFor siRNA experiments, HUVECs were transfected with siRNAs targeting human CD36 (5′-AAGAGGAACTATATTG-TGCCTCCTGTCTC-3′), FATP3 (Santa Cruz Biotechnology, SASI_Hs01_00100092), FATP4 (Santa Cruz Biotechnology, SASI_Hs01_00047-530), ITGβ1 (5′-TGATAGATCCAATGGCTTA-3′), ITGα5 (Bioneer, 1075709), ITGαV (Bioneer, 1075799), ITGβ3 (Bioneer, 1075875), ITGβ5 (Bioneer, 1075906), TIE2 (5′-GGCUAGUAAGAUCAAUGGUdTdT-3′), NFATc1 (5′-CCCGUUCACGUCAGUUUCUACGUCU-3′)or a scrambled control (5′-UAGCGACUAAACACAUCAA-3′) were used.

    Techniques: Transport Assay, Cell Culture, Expressing, Transfection, In Situ, Proximity Ligation Assay, Blocking Assay, Immunoprecipitation, Two Tailed Test

    Angpt2–ITGα5β1 signaling facilitates FA transport through CD36 and FATP3. a , c , d , g – p HUVECs were treated with vehicle or Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) for 15 min or indicated time points. a Comparisons of short-chain FA (BODIPY C-5, 8 μM; n = 4) and long-chain FA (BODIPY C-12 and C-16, 8 μM; n = 4 for vehicle and 3 for Mn 2+ + Angpt2) uptake. b Comparisons of depletion efficiency of CD36 , FATP3 , and FATP4 in corresponding siRNA treated HUVECs. n = 3. c , d Comparisons and representative images of FA (BODIPY C-12, 8 μM) uptake in siControl ( n = 7, 5), si CD36 ( n = 4, 5), si FATP3 ( n = 6), or si FATP4 ( n = 6, 8) HUVECs. Scale bars, 30 μm. e Diagram depicting the endothelial FA transport assay. HUVECs were cultured until confluence on trans-wells and FA (BODIPY C-12, 8 μM) was added to upper layer of trans-well, followed by analysis of transported FA in bottom well. f Comparisons of FA transport for indicated time points after Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) treatment in siControl ( n = 3 for 10, 30 min and 4 for 20 min), si FATP3 ( n = 3) or si FATP4 ( n = 4) HUVECs. n = 3–5. g Comparison of mRNA expression of CD36 and FATP3 . h Representative images of FATP3-td-Tomato transfected HUVECs. Scale bars, 50 μm. i Representative images of CD36 and ITGβ1 expression. Expression of active ITGβ1 (arrows) and CD36 (arrowheads) are co-localized after Angpt2 treatment. Scale bars, 50 μm. j , k Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex. Scale bars, 20 μm. l – n Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex in HUVECs pre-treated with vehicle or ITGα5β1 blocking peptide (ATN-161, 10 μM) for 15 min. Scale bars, 50 μm ( l ); 20 μm ( m ). o , p Immunoprecipitation with anti-IgG and anti-CD36 antibody in HUVECs. Immunoblot analysis with anti-ITGβ1 and anti-ITGα5 antibodies are shown. Graph indicates normalized ratio of immunoprecipitated ITGβ1 per input. Unless otherwise denoted, each dot indicates a mean of triplicate values from three independent experiments and horizontal bars indicate mean ± SD or SEM ( f ) and P values versus control by two-tailed Student’s t test. NS not significant.

    Journal: Nature Communications

    Article Title: Angiopoietin-2–integrin α5β1 signaling enhances vascular fatty acid transport and prevents ectopic lipid-induced insulin resistance

    doi: 10.1038/s41467-020-16795-4

    Figure Lengend Snippet: Angpt2–ITGα5β1 signaling facilitates FA transport through CD36 and FATP3. a , c , d , g – p HUVECs were treated with vehicle or Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) for 15 min or indicated time points. a Comparisons of short-chain FA (BODIPY C-5, 8 μM; n = 4) and long-chain FA (BODIPY C-12 and C-16, 8 μM; n = 4 for vehicle and 3 for Mn 2+ + Angpt2) uptake. b Comparisons of depletion efficiency of CD36 , FATP3 , and FATP4 in corresponding siRNA treated HUVECs. n = 3. c , d Comparisons and representative images of FA (BODIPY C-12, 8 μM) uptake in siControl ( n = 7, 5), si CD36 ( n = 4, 5), si FATP3 ( n = 6), or si FATP4 ( n = 6, 8) HUVECs. Scale bars, 30 μm. e Diagram depicting the endothelial FA transport assay. HUVECs were cultured until confluence on trans-wells and FA (BODIPY C-12, 8 μM) was added to upper layer of trans-well, followed by analysis of transported FA in bottom well. f Comparisons of FA transport for indicated time points after Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) treatment in siControl ( n = 3 for 10, 30 min and 4 for 20 min), si FATP3 ( n = 3) or si FATP4 ( n = 4) HUVECs. n = 3–5. g Comparison of mRNA expression of CD36 and FATP3 . h Representative images of FATP3-td-Tomato transfected HUVECs. Scale bars, 50 μm. i Representative images of CD36 and ITGβ1 expression. Expression of active ITGβ1 (arrows) and CD36 (arrowheads) are co-localized after Angpt2 treatment. Scale bars, 50 μm. j , k Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex. Scale bars, 20 μm. l – n Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex in HUVECs pre-treated with vehicle or ITGα5β1 blocking peptide (ATN-161, 10 μM) for 15 min. Scale bars, 50 μm ( l ); 20 μm ( m ). o , p Immunoprecipitation with anti-IgG and anti-CD36 antibody in HUVECs. Immunoblot analysis with anti-ITGβ1 and anti-ITGα5 antibodies are shown. Graph indicates normalized ratio of immunoprecipitated ITGβ1 per input. Unless otherwise denoted, each dot indicates a mean of triplicate values from three independent experiments and horizontal bars indicate mean ± SD or SEM ( f ) and P values versus control by two-tailed Student’s t test. NS not significant.

    Article Snippet: siRNA transfectionFor siRNA experiments, HUVECs were transfected with siRNAs targeting human CD36 (5′-AAGAGGAACTATATTG-TGCCTCCTGTCTC-3′), FATP3 (Santa Cruz Biotechnology, SASI_Hs01_00100092), FATP4 (Santa Cruz Biotechnology, SASI_Hs01_00047-530), ITGβ1 (5′-TGATAGATCCAATGGCTTA-3′), ITGα5 (Bioneer, 1075709), ITGαV (Bioneer, 1075799), ITGβ3 (Bioneer, 1075875), ITGβ5 (Bioneer, 1075906), TIE2 (5′-GGCUAGUAAGAUCAAUGGUdTdT-3′), NFATc1 (5′-CCCGUUCACGUCAGUUUCUACGUCU-3′)or a scrambled control (5′-UAGCGACUAAACACAUCAA-3′) were used.

    Techniques: Transport Assay, Cell Culture, Expressing, Transfection, In Situ, Proximity Ligation Assay, Blocking Assay, Immunoprecipitation, Two Tailed Test

    ECs of SAT are enriched with integrin α5β1. a Diagram depicting EC-specific RiboTag transcriptome analysis. Inducible recombination of the RiboTag allele led to expression of hemaglutanin (HA)-tagged ribosomal protein (reddish orange) specifically in ECs. Ribosome-bound transcripts were immunoprecipitated (IP) from homogenized whole tissues with anti-HA antibody-coupled magnetic beads. Extracted mRNAs were analyzed by RNA-Seq analysis. b Generation of RiboTag ∆EC mouse, inducible and specific tagging of ribosomes in ECs in 6-week-old mice, and their analyses 2 weeks later. Red arrowheads indicate daily injections of tamoxifen. c Representative images of HA-tagged ribosomal protein in VE-Cadherin expressing ECs in adipose tissues of RiboTag ∆EC mouse. Scale bars, 50 μm. d Comparisons of perilipin and pecam expression in isolated mRNA of ECs from different organs of RiboTag ∆EC mouse. e RNA-seq expression heatmap of ITGα5, ITGβ1, and Tie2 in isolated ECs from different organs using RiboTag ∆EC mouse. n = 5 for each group. f Representative images of ITGα5β1 expression in various adipose tissues of WT mice. Scale bars, 50 μm. g Representative images of Tie2 expression in SAT and BAT of Tie2-GFP mice. Magnified views of dotted-line box are shown in lower panels. Scale bars, 50 μm.

    Journal: Nature Communications

    Article Title: Angiopoietin-2–integrin α5β1 signaling enhances vascular fatty acid transport and prevents ectopic lipid-induced insulin resistance

    doi: 10.1038/s41467-020-16795-4

    Figure Lengend Snippet: ECs of SAT are enriched with integrin α5β1. a Diagram depicting EC-specific RiboTag transcriptome analysis. Inducible recombination of the RiboTag allele led to expression of hemaglutanin (HA)-tagged ribosomal protein (reddish orange) specifically in ECs. Ribosome-bound transcripts were immunoprecipitated (IP) from homogenized whole tissues with anti-HA antibody-coupled magnetic beads. Extracted mRNAs were analyzed by RNA-Seq analysis. b Generation of RiboTag ∆EC mouse, inducible and specific tagging of ribosomes in ECs in 6-week-old mice, and their analyses 2 weeks later. Red arrowheads indicate daily injections of tamoxifen. c Representative images of HA-tagged ribosomal protein in VE-Cadherin expressing ECs in adipose tissues of RiboTag ∆EC mouse. Scale bars, 50 μm. d Comparisons of perilipin and pecam expression in isolated mRNA of ECs from different organs of RiboTag ∆EC mouse. e RNA-seq expression heatmap of ITGα5, ITGβ1, and Tie2 in isolated ECs from different organs using RiboTag ∆EC mouse. n = 5 for each group. f Representative images of ITGα5β1 expression in various adipose tissues of WT mice. Scale bars, 50 μm. g Representative images of Tie2 expression in SAT and BAT of Tie2-GFP mice. Magnified views of dotted-line box are shown in lower panels. Scale bars, 50 μm.

    Article Snippet: siRNA transfectionFor siRNA experiments, HUVECs were transfected with siRNAs targeting human CD36 (5′-AAGAGGAACTATATTG-TGCCTCCTGTCTC-3′), FATP3 (Santa Cruz Biotechnology, SASI_Hs01_00100092), FATP4 (Santa Cruz Biotechnology, SASI_Hs01_00047-530), ITGβ1 (5′-TGATAGATCCAATGGCTTA-3′), ITGα5 (Bioneer, 1075709), ITGαV (Bioneer, 1075799), ITGβ3 (Bioneer, 1075875), ITGβ5 (Bioneer, 1075906), TIE2 (5′-GGCUAGUAAGAUCAAUGGUdTdT-3′), NFATc1 (5′-CCCGUUCACGUCAGUUUCUACGUCU-3′)or a scrambled control (5′-UAGCGACUAAACACAUCAA-3′) were used.

    Techniques: Expressing, Immunoprecipitation, Magnetic Beads, RNA Sequencing Assay, Mouse Assay, Isolation

    Endothelial-to-adipocyte fatty acid transport determines metabolic health. Schematic diagram depicting Angpt2 produced from adipocytes could regulate endothelial FA transport via CD36 and FATP3 through integrin α5β1 signaling to accumulate FAs toward SAT. This process prevents ectopic fat accumulation into endocrine organs, and thus prevents insulin resistance.

    Journal: Nature Communications

    Article Title: Angiopoietin-2–integrin α5β1 signaling enhances vascular fatty acid transport and prevents ectopic lipid-induced insulin resistance

    doi: 10.1038/s41467-020-16795-4

    Figure Lengend Snippet: Endothelial-to-adipocyte fatty acid transport determines metabolic health. Schematic diagram depicting Angpt2 produced from adipocytes could regulate endothelial FA transport via CD36 and FATP3 through integrin α5β1 signaling to accumulate FAs toward SAT. This process prevents ectopic fat accumulation into endocrine organs, and thus prevents insulin resistance.

    Article Snippet: siRNA transfectionFor siRNA experiments, HUVECs were transfected with siRNAs targeting human CD36 (5′-AAGAGGAACTATATTG-TGCCTCCTGTCTC-3′), FATP3 (Santa Cruz Biotechnology, SASI_Hs01_00100092), FATP4 (Santa Cruz Biotechnology, SASI_Hs01_00047-530), ITGβ1 (5′-TGATAGATCCAATGGCTTA-3′), ITGα5 (Bioneer, 1075709), ITGαV (Bioneer, 1075799), ITGβ3 (Bioneer, 1075875), ITGβ5 (Bioneer, 1075906), TIE2 (5′-GGCUAGUAAGAUCAAUGGUdTdT-3′), NFATc1 (5′-CCCGUUCACGUCAGUUUCUACGUCU-3′)or a scrambled control (5′-UAGCGACUAAACACAUCAA-3′) were used.

    Techniques: Produced

    Angpt2–ITGα5β1 signaling facilitates FA transport through CD36 and FATP3. a , c , d , g – p HUVECs were treated with vehicle or Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) for 15 min or indicated time points. a Comparisons of short-chain FA (BODIPY C-5, 8 μM; n = 4) and long-chain FA (BODIPY C-12 and C-16, 8 μM; n = 4 for vehicle and 3 for Mn 2+ + Angpt2) uptake. b Comparisons of depletion efficiency of CD36 , FATP3 , and FATP4 in corresponding siRNA treated HUVECs. n = 3. c , d Comparisons and representative images of FA (BODIPY C-12, 8 μM) uptake in siControl ( n = 7, 5), si CD36 ( n = 4, 5), si FATP3 ( n = 6), or si FATP4 ( n = 6, 8) HUVECs. Scale bars, 30 μm. e Diagram depicting the endothelial FA transport assay. HUVECs were cultured until confluence on trans-wells and FA (BODIPY C-12, 8 μM) was added to upper layer of trans-well, followed by analysis of transported FA in bottom well. f Comparisons of FA transport for indicated time points after Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) treatment in siControl ( n = 3 for 10, 30 min and 4 for 20 min), si FATP3 ( n = 3) or si FATP4 ( n = 4) HUVECs. n = 3–5. g Comparison of mRNA expression of CD36 and FATP3 . h Representative images of FATP3-td-Tomato transfected HUVECs. Scale bars, 50 μm. i Representative images of CD36 and ITGβ1 expression. Expression of active ITGβ1 (arrows) and CD36 (arrowheads) are co-localized after Angpt2 treatment. Scale bars, 50 μm. j , k Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex. Scale bars, 20 μm. l – n Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex in HUVECs pre-treated with vehicle or ITGα5β1 blocking peptide (ATN-161, 10 μM) for 15 min. Scale bars, 50 μm ( l ); 20 μm ( m ). o , p Immunoprecipitation with anti-IgG and anti-CD36 antibody in HUVECs. Immunoblot analysis with anti-ITGβ1 and anti-ITGα5 antibodies are shown. Graph indicates normalized ratio of immunoprecipitated ITGβ1 per input. Unless otherwise denoted, each dot indicates a mean of triplicate values from three independent experiments and horizontal bars indicate mean ± SD or SEM ( f ) and P values versus control by two-tailed Student’s t test. NS not significant.

    Journal: Nature Communications

    Article Title: Angiopoietin-2–integrin α5β1 signaling enhances vascular fatty acid transport and prevents ectopic lipid-induced insulin resistance

    doi: 10.1038/s41467-020-16795-4

    Figure Lengend Snippet: Angpt2–ITGα5β1 signaling facilitates FA transport through CD36 and FATP3. a , c , d , g – p HUVECs were treated with vehicle or Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) for 15 min or indicated time points. a Comparisons of short-chain FA (BODIPY C-5, 8 μM; n = 4) and long-chain FA (BODIPY C-12 and C-16, 8 μM; n = 4 for vehicle and 3 for Mn 2+ + Angpt2) uptake. b Comparisons of depletion efficiency of CD36 , FATP3 , and FATP4 in corresponding siRNA treated HUVECs. n = 3. c , d Comparisons and representative images of FA (BODIPY C-12, 8 μM) uptake in siControl ( n = 7, 5), si CD36 ( n = 4, 5), si FATP3 ( n = 6), or si FATP4 ( n = 6, 8) HUVECs. Scale bars, 30 μm. e Diagram depicting the endothelial FA transport assay. HUVECs were cultured until confluence on trans-wells and FA (BODIPY C-12, 8 μM) was added to upper layer of trans-well, followed by analysis of transported FA in bottom well. f Comparisons of FA transport for indicated time points after Mn 2+ (1 mM) + Angpt2 (2.5 μg/ml) treatment in siControl ( n = 3 for 10, 30 min and 4 for 20 min), si FATP3 ( n = 3) or si FATP4 ( n = 4) HUVECs. n = 3–5. g Comparison of mRNA expression of CD36 and FATP3 . h Representative images of FATP3-td-Tomato transfected HUVECs. Scale bars, 50 μm. i Representative images of CD36 and ITGβ1 expression. Expression of active ITGβ1 (arrows) and CD36 (arrowheads) are co-localized after Angpt2 treatment. Scale bars, 50 μm. j , k Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex. Scale bars, 20 μm. l – n Representative images and comparison of in situ proximity ligation assay between CD36 and ITGβ1 complex in HUVECs pre-treated with vehicle or ITGα5β1 blocking peptide (ATN-161, 10 μM) for 15 min. Scale bars, 50 μm ( l ); 20 μm ( m ). o , p Immunoprecipitation with anti-IgG and anti-CD36 antibody in HUVECs. Immunoblot analysis with anti-ITGβ1 and anti-ITGα5 antibodies are shown. Graph indicates normalized ratio of immunoprecipitated ITGβ1 per input. Unless otherwise denoted, each dot indicates a mean of triplicate values from three independent experiments and horizontal bars indicate mean ± SD or SEM ( f ) and P values versus control by two-tailed Student’s t test. NS not significant.

    Article Snippet: siRNA transfectionFor siRNA experiments, HUVECs were transfected with siRNAs targeting human CD36 (5′-AAGAGGAACTATATTG-TGCCTCCTGTCTC-3′), FATP3 (Santa Cruz Biotechnology, SASI_Hs01_00100092), FATP4 (Santa Cruz Biotechnology, SASI_Hs01_00047-530), ITGβ1 (5′-TGATAGATCCAATGGCTTA-3′), ITGα5 (Bioneer, 1075709), ITGαV (Bioneer, 1075799), ITGβ3 (Bioneer, 1075875), ITGβ5 (Bioneer, 1075906), TIE2 (5′-GGCUAGUAAGAUCAAUGGUdTdT-3′), NFATc1 (5′-CCCGUUCACGUCAGUUUCUACGUCU-3′)or a scrambled control (5′-UAGCGACUAAACACAUCAA-3′) were used.

    Techniques: Transport Assay, Cell Culture, Expressing, Transfection, In Situ, Proximity Ligation Assay, Blocking Assay, Immunoprecipitation, Two Tailed Test

    Knockdown of HIF-1α blocks dexrazoxane (DRZ)-mediated cardioprotection. (A) Transfection efficiency in H9c2 cells. The figure shows a merged image with DAPI-stained nuclei (blue) and punctuate fluorescence of rhodamine-labelled siRNA (red). (B)

    Journal: British Journal of Pharmacology

    Article Title: Role of hypoxia-inducible factors in the dexrazoxane-mediated protection of cardiomyocytes from doxorubicin-induced toxicity

    doi: 10.1111/j.1476-5381.2011.01208.x

    Figure Lengend Snippet: Knockdown of HIF-1α blocks dexrazoxane (DRZ)-mediated cardioprotection. (A) Transfection efficiency in H9c2 cells. The figure shows a merged image with DAPI-stained nuclei (blue) and punctuate fluorescence of rhodamine-labelled siRNA (red). (B)

    Article Snippet: To monitor transfection efficiency, the cells were transfected with a rhodamine-labelled siRNA (Qiagen SpA, Milano, Italy), fixed and observed using fluorescence microscopy (excitation 530 nm, emission 570 nm); nuclei were counterstained with 10 µg·mL−1 4′-6-diamidino-2-phenylindole (DAPI, Sigma), and fluorescence was observed (excitation 364 nm, emission 454 nm).

    Techniques: Transfection, Staining, Fluorescence

    Analysis of AAT Expression after Transfection with Liposomes or Lipofectamine 2000 Containing AAT-Encoding mRNA Concentration of AAT protein in cell supernatants after transfection with AAT mRNA encapsulated in liposomes or Lipofectamine 2000 for 24 or 72 hr. Data are shown as mean ± SEM (n = 5). **p

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: Cationic Nanoliposomes Meet mRNA: Efficient Delivery of Modified mRNA Using Hemocompatible and Stable Vectors for Therapeutic Applications

    doi: 10.1016/j.omtn.2017.07.013

    Figure Lengend Snippet: Analysis of AAT Expression after Transfection with Liposomes or Lipofectamine 2000 Containing AAT-Encoding mRNA Concentration of AAT protein in cell supernatants after transfection with AAT mRNA encapsulated in liposomes or Lipofectamine 2000 for 24 or 72 hr. Data are shown as mean ± SEM (n = 5). **p

    Article Snippet: Additionally, lipoplexes were formed by mixing 1 μg of EGFP mRNA and liposomes or Lipofectamine 2000 (Invitrogen) and incubated for 20 min at RT.

    Techniques: Expressing, Transfection, Concentration Assay

    Preparation and Characterization of Liposomes (A) Schematic overview of the liposome production process including lipid preparation and mixing, drying under O 2 -free condition, and evaporation overnight in vacuum. The developed lipid cake was rehydrated with water followed by sonification, extrusion, and homogenization to get unilamellar lipid vesicles. (B) Visualization of the liposomes using TEM after negative staining. (C) Analysis of the mRNA encapsulation efficacy of liposomes compared to Lipofectamine 2000. Data are shown as mean ± SEM (n = 5).

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: Cationic Nanoliposomes Meet mRNA: Efficient Delivery of Modified mRNA Using Hemocompatible and Stable Vectors for Therapeutic Applications

    doi: 10.1016/j.omtn.2017.07.013

    Figure Lengend Snippet: Preparation and Characterization of Liposomes (A) Schematic overview of the liposome production process including lipid preparation and mixing, drying under O 2 -free condition, and evaporation overnight in vacuum. The developed lipid cake was rehydrated with water followed by sonification, extrusion, and homogenization to get unilamellar lipid vesicles. (B) Visualization of the liposomes using TEM after negative staining. (C) Analysis of the mRNA encapsulation efficacy of liposomes compared to Lipofectamine 2000. Data are shown as mean ± SEM (n = 5).

    Article Snippet: Additionally, lipoplexes were formed by mixing 1 μg of EGFP mRNA and liposomes or Lipofectamine 2000 (Invitrogen) and incubated for 20 min at RT.

    Techniques: Evaporation, Homogenization, Transmission Electron Microscopy, Negative Staining

    MRPL10 knockdown leads to decreased mitochondrial activity and mitochondrial complex expression. (A) Several components of mitochondrial respiratory complexes from mitochondria lysates in HEK293 cells with or without MRPL10 knockdown were subjected to SDS-PAGE and immunoblotted with the indicated antibodies. Bar graphs depict the signal intensities of protein indicated. The relative expression is shown as a percentage of the mean using the control sample as reference. (B) Activities of the five mitochondrial respiratory complexes were measured in mitochondria isolated from control or MRPL10 knockdown HEK293 cells. Cells treated with rotenone (5 μM), TTFA (10 μM), antimycin A (2 μM), KCN (100 μM), and oligomycin (10 μM) to inhibit mitochondrial complex I, II, III, IV, and V activities were used as positive controls. (C) Mitchondrial respiration rate was measured using a clark type electrode. Cells treated with mitochondrial complex I inhibitor rotenone (5 μM) were used as positive controls. (D) Comparison of the ATP production in control or MRPL10 knockdown cells. Cells treated with mitochondrial complex I inhibitor rotenone (5 μM) were used as positive controls. (E) The copy number of mtDNA was measured by quantitative PCR analysis. (F) Mitochondrial membrane potential was detected by FACS analysis in control or MRPL10 knockdown HEK293 cells by three independent experiments. Cells treated with CCCP (10 μM) for 2 h were used as positive controls. The results were obtained from three independent experiments. The relative expression is shown as a percentage of the mean using the control sample as reference. All data in this figure represent the mean ± SEM of three independent experiments. * p

    Journal: DNA and Cell Biology

    Article Title: Mitochondrial Ribosomal Protein L10 Associates with Cyclin B1/Cdk1 Activity and Mitochondrial Function

    doi: 10.1089/dna.2016.3271

    Figure Lengend Snippet: MRPL10 knockdown leads to decreased mitochondrial activity and mitochondrial complex expression. (A) Several components of mitochondrial respiratory complexes from mitochondria lysates in HEK293 cells with or without MRPL10 knockdown were subjected to SDS-PAGE and immunoblotted with the indicated antibodies. Bar graphs depict the signal intensities of protein indicated. The relative expression is shown as a percentage of the mean using the control sample as reference. (B) Activities of the five mitochondrial respiratory complexes were measured in mitochondria isolated from control or MRPL10 knockdown HEK293 cells. Cells treated with rotenone (5 μM), TTFA (10 μM), antimycin A (2 μM), KCN (100 μM), and oligomycin (10 μM) to inhibit mitochondrial complex I, II, III, IV, and V activities were used as positive controls. (C) Mitchondrial respiration rate was measured using a clark type electrode. Cells treated with mitochondrial complex I inhibitor rotenone (5 μM) were used as positive controls. (D) Comparison of the ATP production in control or MRPL10 knockdown cells. Cells treated with mitochondrial complex I inhibitor rotenone (5 μM) were used as positive controls. (E) The copy number of mtDNA was measured by quantitative PCR analysis. (F) Mitochondrial membrane potential was detected by FACS analysis in control or MRPL10 knockdown HEK293 cells by three independent experiments. Cells treated with CCCP (10 μM) for 2 h were used as positive controls. The results were obtained from three independent experiments. The relative expression is shown as a percentage of the mean using the control sample as reference. All data in this figure represent the mean ± SEM of three independent experiments. * p

    Article Snippet: Briefly, HEK293 cells were transfected with MRPL10 siRNAs twice for 72 h followed by staining with TMRE (100 nM) for 20 min. Post treatment, cells were immediately analyzed using a FACScan flow cytometer (Becton Dickinson, San Jose, CA) with excitation at 488 nm and emission at 530 nm.

    Techniques: Activity Assay, Expressing, SDS Page, Isolation, Real-time Polymerase Chain Reaction, FACS

    Cyclin B restores mitochondrial abnormality caused by MRPL10 knockdown. (A, B) HEK293 cells were transfected with MRPL10 siRNA and Cyclin B1 in different combination. Activities of the mitochondrial respiratory complexes were measured in isolated mitochondria. (C) Mitchondrial respiration rate was measured using a clark type electrode in each group. (D) Comparison of the ATP production in each group. (E) HEK293 cells were transfected with MRPL10 siRNA and Cyclin B1 in different combination for 72 h, mitochondrial proteins were separated and immunoprecipitated using a phosphoserine/threonine antibody followed by immunoblotting using antibodies to NDUFA12 and NDUFB6. Direct immunoblotting of cell lysates with the indicated antibody are shown as the input control ( left panel ). β-actin is shown as loading control. All data in this figure represent the mean ± SEM of three independent experiments. * p

    Journal: DNA and Cell Biology

    Article Title: Mitochondrial Ribosomal Protein L10 Associates with Cyclin B1/Cdk1 Activity and Mitochondrial Function

    doi: 10.1089/dna.2016.3271

    Figure Lengend Snippet: Cyclin B restores mitochondrial abnormality caused by MRPL10 knockdown. (A, B) HEK293 cells were transfected with MRPL10 siRNA and Cyclin B1 in different combination. Activities of the mitochondrial respiratory complexes were measured in isolated mitochondria. (C) Mitchondrial respiration rate was measured using a clark type electrode in each group. (D) Comparison of the ATP production in each group. (E) HEK293 cells were transfected with MRPL10 siRNA and Cyclin B1 in different combination for 72 h, mitochondrial proteins were separated and immunoprecipitated using a phosphoserine/threonine antibody followed by immunoblotting using antibodies to NDUFA12 and NDUFB6. Direct immunoblotting of cell lysates with the indicated antibody are shown as the input control ( left panel ). β-actin is shown as loading control. All data in this figure represent the mean ± SEM of three independent experiments. * p

    Article Snippet: Briefly, HEK293 cells were transfected with MRPL10 siRNAs twice for 72 h followed by staining with TMRE (100 nM) for 20 min. Post treatment, cells were immediately analyzed using a FACScan flow cytometer (Becton Dickinson, San Jose, CA) with excitation at 488 nm and emission at 530 nm.

    Techniques: Transfection, Isolation, Immunoprecipitation