anti-cd49d Search Results


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  • 94
    Bio X Cell anti vla 4
    Assessing the relative role of self antigens and microbiota in T cell residence in SLOs. a – c 6–12-week-old C57BL/6 Foxp3-GFP mice were injected or not i.p. with 200 μg of anti-LFA-1 (αL) and <t>anti-VLA-4</t> (α4) Abs. Forty-eight hours later, peripheral lymph nodes (pLN) were harvested and analyzed. CD5 ( a ) and Nur77 ( b ) fluorescence histograms of CD4 Treg and CD4 Tmem cells from the pLNs of a representative treated and a representative control C57BL/6 Foxp3-GFP mouse. Quantification is shown as means ± SEM with unpaired t -test on the right part of these panels. c – f 6–8-week-old SPF or germ-free C57BL/6 mice were injected or not i.p. with 200 μg of anti-LFA-1 (αL) and anti-VLA-4 (α4) Abs. Forty-eight hours later, SLOs were harvested and analyzed. c Diagram illustrating the experimental model. d Total cell numbers recovered from pLNs, mLNs, and Peyer’s patches (Pp) of SPF or germ-free mice. e Recovery of CD4 Treg and CD4 Tmem cells in pLNs and mLNs. f Recovery of CD4 Treg, CD4 Tmem and CD8 Tmem cells in Peyer’s patches (Pp). Each dot represents an individual mouse (unpaired t -test). * p
    Anti Vla 4, supplied by Bio X Cell, used in various techniques. Bioz Stars score: 94/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher anti cd49d
    Assessing the relative role of self antigens and microbiota in T cell residence in SLOs. a – c 6–12-week-old C57BL/6 Foxp3-GFP mice were injected or not i.p. with 200 μg of anti-LFA-1 (αL) and <t>anti-VLA-4</t> (α4) Abs. Forty-eight hours later, peripheral lymph nodes (pLN) were harvested and analyzed. CD5 ( a ) and Nur77 ( b ) fluorescence histograms of CD4 Treg and CD4 Tmem cells from the pLNs of a representative treated and a representative control C57BL/6 Foxp3-GFP mouse. Quantification is shown as means ± SEM with unpaired t -test on the right part of these panels. c – f 6–8-week-old SPF or germ-free C57BL/6 mice were injected or not i.p. with 200 μg of anti-LFA-1 (αL) and anti-VLA-4 (α4) Abs. Forty-eight hours later, SLOs were harvested and analyzed. c Diagram illustrating the experimental model. d Total cell numbers recovered from pLNs, mLNs, and Peyer’s patches (Pp) of SPF or germ-free mice. e Recovery of CD4 Treg and CD4 Tmem cells in pLNs and mLNs. f Recovery of CD4 Treg, CD4 Tmem and CD8 Tmem cells in Peyer’s patches (Pp). Each dot represents an individual mouse (unpaired t -test). * p
    Anti Cd49d, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 211 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Becton Dickinson anti cd49d
    TGF-β suppression of EOMES-driven responses was common to CD4 T cells from mice infected with MCMV and HIV-infected patients. ( A and B ) Mixed chimeras with 1:1 ratio of WT (CD45.1, black) and ERCre + Tgfbr2 flox (CD45.2, red) bone marrow reconstituted prior to TGFβ-RII deletion, were infected with 2 × 10 4 PFU MCMV i.p. ( A ) Proportion of CD4 T cells expressing activation markers CD11a and <t>CD49d</t> over time after gating on congenic marker, is shown for postinfection day 14. ( B ) Overlays of EOMES and KLRG expression in activated CD4 T cells. ( C ) Upper panels show density plots of mRNA expression level vs. CD107 protein and quantification of Eomes and GrzB transcripts in CD107 + vs. CD107 – responding CD4 T cells. Lower panels show single-cell analysis of Eomes and granzyme B mRNA expression levels in HIV-specific IFN-γ + CD107a + CD4 T cells after a 5-hour Gag peptide stimulation using Biomark Fluidigm analysis. The transcript expression threshold (Et) was defined as the qPCR cycle number above background at which the transcript was detected. ( D and E ) Gag-responsive HIV-specific CD4 T cells were isolated and cultured for 5 days in the presence or absence of TGF-β and restimulated with GAG peptides in the presence of CD28/CD49d, and monensin for percentage of CD107 + CD4 T cells ( D ) and eomesodermin and granzyme B expression in HIV-specific CD107 + CD4 T cells ( E ). ( A and B ) Representative data from 2 independent experiments, with n = 4 or 5 mice/group. ( C – E ) Representative data from n = 10 HIV + treatment-naive subjects. Two-way ANOVA ( A ), paired t test ( B – E ), * P
    Anti Cd49d, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 2496 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    BioLegend anti cd49d
    TGF-β suppression of EOMES-driven responses was common to CD4 T cells from mice infected with MCMV and HIV-infected patients. ( A and B ) Mixed chimeras with 1:1 ratio of WT (CD45.1, black) and ERCre + Tgfbr2 flox (CD45.2, red) bone marrow reconstituted prior to TGFβ-RII deletion, were infected with 2 × 10 4 PFU MCMV i.p. ( A ) Proportion of CD4 T cells expressing activation markers CD11a and <t>CD49d</t> over time after gating on congenic marker, is shown for postinfection day 14. ( B ) Overlays of EOMES and KLRG expression in activated CD4 T cells. ( C ) Upper panels show density plots of mRNA expression level vs. CD107 protein and quantification of Eomes and GrzB transcripts in CD107 + vs. CD107 – responding CD4 T cells. Lower panels show single-cell analysis of Eomes and granzyme B mRNA expression levels in HIV-specific IFN-γ + CD107a + CD4 T cells after a 5-hour Gag peptide stimulation using Biomark Fluidigm analysis. The transcript expression threshold (Et) was defined as the qPCR cycle number above background at which the transcript was detected. ( D and E ) Gag-responsive HIV-specific CD4 T cells were isolated and cultured for 5 days in the presence or absence of TGF-β and restimulated with GAG peptides in the presence of CD28/CD49d, and monensin for percentage of CD107 + CD4 T cells ( D ) and eomesodermin and granzyme B expression in HIV-specific CD107 + CD4 T cells ( E ). ( A and B ) Representative data from 2 independent experiments, with n = 4 or 5 mice/group. ( C – E ) Representative data from n = 10 HIV + treatment-naive subjects. Two-way ANOVA ( A ), paired t test ( B – E ), * P
    Anti Cd49d, supplied by BioLegend, used in various techniques. Bioz Stars score: 94/100, based on 163 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Millipore anti cd49d
    TGF-β suppression of EOMES-driven responses was common to CD4 T cells from mice infected with MCMV and HIV-infected patients. ( A and B ) Mixed chimeras with 1:1 ratio of WT (CD45.1, black) and ERCre + Tgfbr2 flox (CD45.2, red) bone marrow reconstituted prior to TGFβ-RII deletion, were infected with 2 × 10 4 PFU MCMV i.p. ( A ) Proportion of CD4 T cells expressing activation markers CD11a and <t>CD49d</t> over time after gating on congenic marker, is shown for postinfection day 14. ( B ) Overlays of EOMES and KLRG expression in activated CD4 T cells. ( C ) Upper panels show density plots of mRNA expression level vs. CD107 protein and quantification of Eomes and GrzB transcripts in CD107 + vs. CD107 – responding CD4 T cells. Lower panels show single-cell analysis of Eomes and granzyme B mRNA expression levels in HIV-specific IFN-γ + CD107a + CD4 T cells after a 5-hour Gag peptide stimulation using Biomark Fluidigm analysis. The transcript expression threshold (Et) was defined as the qPCR cycle number above background at which the transcript was detected. ( D and E ) Gag-responsive HIV-specific CD4 T cells were isolated and cultured for 5 days in the presence or absence of TGF-β and restimulated with GAG peptides in the presence of CD28/CD49d, and monensin for percentage of CD107 + CD4 T cells ( D ) and eomesodermin and granzyme B expression in HIV-specific CD107 + CD4 T cells ( E ). ( A and B ) Representative data from 2 independent experiments, with n = 4 or 5 mice/group. ( C – E ) Representative data from n = 10 HIV + treatment-naive subjects. Two-way ANOVA ( A ), paired t test ( B – E ), * P
    Anti Cd49d, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 86 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Pharmingen anti cd49d
    Expression of CXCR4 or <t>CD49d</t> (VLA-4) by different B-cell lines or normal blood B cells, as indicated. Displayed are fluorescence histograms depicting the relative red fluorescence intensity of the cells stained with anti-CXCR4 mAb’s ( a , shaded histograms) or with anti-CD49d mAb’s ( b , shaded histograms) compared with that of the same cells stained with a PE-conjugated isotype control mAb of irrelevant specificity (open histograms). The mean fluorescence intensity ratio of each specifically-stained cell population is displayed above the histograms.
    Anti Cd49d, supplied by Pharmingen, used in various techniques. Bioz Stars score: 92/100, based on 81 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    BioLegend apc anti human cd49d
    Expression of CXCR4 or <t>CD49d</t> (VLA-4) by different B-cell lines or normal blood B cells, as indicated. Displayed are fluorescence histograms depicting the relative red fluorescence intensity of the cells stained with anti-CXCR4 mAb’s ( a , shaded histograms) or with anti-CD49d mAb’s ( b , shaded histograms) compared with that of the same cells stained with a PE-conjugated isotype control mAb of irrelevant specificity (open histograms). The mean fluorescence intensity ratio of each specifically-stained cell population is displayed above the histograms.
    Apc Anti Human Cd49d, supplied by BioLegend, used in various techniques. Bioz Stars score: 94/100, based on 43 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Bio-Rad anti cd49d
    The percentages of monocytes positive in FACS analysis for ‘fibronectin receptors’ (FN-FITC, see text), CD49e, <t>CD49d,</t> CD29, CD49b and CD49a integrins. Mean ± s.d. are given of original monocytes (first two columns of a set; □ healthy controls, ▪ patients) and of FN-adhered monocytes (last two columns of a set; healthy controls, patients) of healthy controls and patients. a = P
    Anti Cd49d, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 91/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    BioLegend pe cy7 anti human cd49d
    The percentages of monocytes positive in FACS analysis for ‘fibronectin receptors’ (FN-FITC, see text), CD49e, <t>CD49d,</t> CD29, CD49b and CD49a integrins. Mean ± s.d. are given of original monocytes (first two columns of a set; □ healthy controls, ▪ patients) and of FN-adhered monocytes (last two columns of a set; healthy controls, patients) of healthy controls and patients. a = P
    Pe Cy7 Anti Human Cd49d, supplied by BioLegend, used in various techniques. Bioz Stars score: 95/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Becton Dickinson anti cd28 cd49d co stimulus
    The percentages of monocytes positive in FACS analysis for ‘fibronectin receptors’ (FN-FITC, see text), CD49e, <t>CD49d,</t> CD29, CD49b and CD49a integrins. Mean ± s.d. are given of original monocytes (first two columns of a set; □ healthy controls, ▪ patients) and of FN-adhered monocytes (last two columns of a set; healthy controls, patients) of healthy controls and patients. a = P
    Anti Cd28 Cd49d Co Stimulus, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 91/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Thermo Fisher anti cd49d antibodies
    The percentages of monocytes positive in FACS analysis for ‘fibronectin receptors’ (FN-FITC, see text), CD49e, <t>CD49d,</t> CD29, CD49b and CD49a integrins. Mean ± s.d. are given of original monocytes (first two columns of a set; □ healthy controls, ▪ patients) and of FN-adhered monocytes (last two columns of a set; healthy controls, patients) of healthy controls and patients. a = P
    Anti Cd49d Antibodies, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    BioLegend anti cd49d fitc
    Characterization of MSCs derived from rat bone marrow and transfection efficiency of MSCs with pEF1/His C::IGF-1::EGFP. (A) Expression levels of surface markers in the isolated MSCs were measured by flow cytometry for <t>CD29-FITC,</t> CD54-PE, CD90-FITC, CD45-PE and <t>CD49d-FITC.</t> The red histograms represent staining with respective surface marker antibodies and the black histograms represent staining with isotype controls. (B) Prior to FACS, the transfection efficiency of MSCs with pEF1/His C::IGF-1::EGFP and the nonviral carrier PAM-ABP was ~20% (P4). (C) Following FACS, transfected MSCs were collected and the purity was increased to 95.1%. Therefore, approximately all MSCs expressed a green fluorescent signal and IGF-1. CD, cluster of differentiation; EGFP, enhanced green fluorescence protein; IGF, insulin-like growth factor; MSC, mesenchymal stem cells; FACS, fluorescence-activated cell sorting cell sorting; FITC, fluorescein isothiocyanate; PE, phycoerythrin.
    Anti Cd49d Fitc, supplied by BioLegend, used in various techniques. Bioz Stars score: 93/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    BioLegend anti human cd49d
    Characterization of MSCs derived from rat bone marrow and transfection efficiency of MSCs with pEF1/His C::IGF-1::EGFP. (A) Expression levels of surface markers in the isolated MSCs were measured by flow cytometry for <t>CD29-FITC,</t> CD54-PE, CD90-FITC, CD45-PE and <t>CD49d-FITC.</t> The red histograms represent staining with respective surface marker antibodies and the black histograms represent staining with isotype controls. (B) Prior to FACS, the transfection efficiency of MSCs with pEF1/His C::IGF-1::EGFP and the nonviral carrier PAM-ABP was ~20% (P4). (C) Following FACS, transfected MSCs were collected and the purity was increased to 95.1%. Therefore, approximately all MSCs expressed a green fluorescent signal and IGF-1. CD, cluster of differentiation; EGFP, enhanced green fluorescence protein; IGF, insulin-like growth factor; MSC, mesenchymal stem cells; FACS, fluorescence-activated cell sorting cell sorting; FITC, fluorescein isothiocyanate; PE, phycoerythrin.
    Anti Human Cd49d, supplied by BioLegend, used in various techniques. Bioz Stars score: 94/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher anti α4 integrin
    Cytofluorimetric characterization of KMSC (A) and KMSC-derived MPs (B). One µm and 6 µm beads were used as internal size standards. P1 area was compatible with 1 µm. Red lines indicate the expression of annexin V, CD44, CD73, CD29, <t>α4</t> <t>integrin,</t> α5 integrin, and α6 integrin surface molecules. Black lines indicate isotypic controls.
    Anti α4 Integrin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Assessing the relative role of self antigens and microbiota in T cell residence in SLOs. a – c 6–12-week-old C57BL/6 Foxp3-GFP mice were injected or not i.p. with 200 μg of anti-LFA-1 (αL) and anti-VLA-4 (α4) Abs. Forty-eight hours later, peripheral lymph nodes (pLN) were harvested and analyzed. CD5 ( a ) and Nur77 ( b ) fluorescence histograms of CD4 Treg and CD4 Tmem cells from the pLNs of a representative treated and a representative control C57BL/6 Foxp3-GFP mouse. Quantification is shown as means ± SEM with unpaired t -test on the right part of these panels. c – f 6–8-week-old SPF or germ-free C57BL/6 mice were injected or not i.p. with 200 μg of anti-LFA-1 (αL) and anti-VLA-4 (α4) Abs. Forty-eight hours later, SLOs were harvested and analyzed. c Diagram illustrating the experimental model. d Total cell numbers recovered from pLNs, mLNs, and Peyer’s patches (Pp) of SPF or germ-free mice. e Recovery of CD4 Treg and CD4 Tmem cells in pLNs and mLNs. f Recovery of CD4 Treg, CD4 Tmem and CD8 Tmem cells in Peyer’s patches (Pp). Each dot represents an individual mouse (unpaired t -test). * p

    Journal: Nature Communications

    Article Title: Profiling the lymphoid-resident T cell pool reveals modulation by age and microbiota

    doi: 10.1038/s41467-017-02458-4

    Figure Lengend Snippet: Assessing the relative role of self antigens and microbiota in T cell residence in SLOs. a – c 6–12-week-old C57BL/6 Foxp3-GFP mice were injected or not i.p. with 200 μg of anti-LFA-1 (αL) and anti-VLA-4 (α4) Abs. Forty-eight hours later, peripheral lymph nodes (pLN) were harvested and analyzed. CD5 ( a ) and Nur77 ( b ) fluorescence histograms of CD4 Treg and CD4 Tmem cells from the pLNs of a representative treated and a representative control C57BL/6 Foxp3-GFP mouse. Quantification is shown as means ± SEM with unpaired t -test on the right part of these panels. c – f 6–8-week-old SPF or germ-free C57BL/6 mice were injected or not i.p. with 200 μg of anti-LFA-1 (αL) and anti-VLA-4 (α4) Abs. Forty-eight hours later, SLOs were harvested and analyzed. c Diagram illustrating the experimental model. d Total cell numbers recovered from pLNs, mLNs, and Peyer’s patches (Pp) of SPF or germ-free mice. e Recovery of CD4 Treg and CD4 Tmem cells in pLNs and mLNs. f Recovery of CD4 Treg, CD4 Tmem and CD8 Tmem cells in Peyer’s patches (Pp). Each dot represents an individual mouse (unpaired t -test). * p

    Article Snippet: Blocking T cell entry into LNs and Pp C57BL/6 Foxp3-GFP mice were injected or not i.p. with 200 μg of anti-LFA-1 (clone M17/4) and anti-VLA-4 (clone PS/2) Abs (BioXcell) to block T cell entry into LNs and Peyer’s patches as previously described .

    Techniques: Mouse Assay, Injection, Fluorescence

    Analysis of residual T cells after blocking T cell entry into LNs and Peyer’s Patches. a – c 6–12-week-old C57BL/6 Foxp3-GFP mice were injected or not i.p. with 200 μg of anti-LFA-1 (αL) and anti-VLA-4 (α4) Abs. Twenty-four hours later, cLNs, mLNs, Pp, and spleen were harvested and analyzed. a Diagram illustrating the experimental model. b Distribution of Treg, Tmem, and Tnaive cells among CD4 T cells in the indicated SLOs of treated or untreated mice. c Distribution of Treg, Tmem, and Tnaive cells among CD8 T cells in the indicated SLOs of treated or untreated mice. d , e 6–12-week-old C57BL/6 Foxp3-GFP mice were injected or not i.p. with 200 μg of anti-LFA-1 (αL) and anti-VLA-4 (α4) Abs every 2 days from day 0 to day 6 and SLOs were recovered for analysis at various time-points. d Experimental model. e Percentages of Treg, Tmem, and Tnaive cells among CD4 T cells are shown as means ± SEM for the indicated SLOs. f Relative absolute numbers of CD4 Treg, Tmem, and Tnaive cells in cLNs, mLNs, and spleen are shown. The percentage of recovery was calculated by dividing the absolute numbers in treated mice by the mean absolute number obtained in untreated animals. Data are means ± SEM for at least three independent experiments. Mouse clip arts were generated in ref. 21

    Journal: Nature Communications

    Article Title: Profiling the lymphoid-resident T cell pool reveals modulation by age and microbiota

    doi: 10.1038/s41467-017-02458-4

    Figure Lengend Snippet: Analysis of residual T cells after blocking T cell entry into LNs and Peyer’s Patches. a – c 6–12-week-old C57BL/6 Foxp3-GFP mice were injected or not i.p. with 200 μg of anti-LFA-1 (αL) and anti-VLA-4 (α4) Abs. Twenty-four hours later, cLNs, mLNs, Pp, and spleen were harvested and analyzed. a Diagram illustrating the experimental model. b Distribution of Treg, Tmem, and Tnaive cells among CD4 T cells in the indicated SLOs of treated or untreated mice. c Distribution of Treg, Tmem, and Tnaive cells among CD8 T cells in the indicated SLOs of treated or untreated mice. d , e 6–12-week-old C57BL/6 Foxp3-GFP mice were injected or not i.p. with 200 μg of anti-LFA-1 (αL) and anti-VLA-4 (α4) Abs every 2 days from day 0 to day 6 and SLOs were recovered for analysis at various time-points. d Experimental model. e Percentages of Treg, Tmem, and Tnaive cells among CD4 T cells are shown as means ± SEM for the indicated SLOs. f Relative absolute numbers of CD4 Treg, Tmem, and Tnaive cells in cLNs, mLNs, and spleen are shown. The percentage of recovery was calculated by dividing the absolute numbers in treated mice by the mean absolute number obtained in untreated animals. Data are means ± SEM for at least three independent experiments. Mouse clip arts were generated in ref. 21

    Article Snippet: Blocking T cell entry into LNs and Pp C57BL/6 Foxp3-GFP mice were injected or not i.p. with 200 μg of anti-LFA-1 (clone M17/4) and anti-VLA-4 (clone PS/2) Abs (BioXcell) to block T cell entry into LNs and Peyer’s patches as previously described .

    Techniques: Blocking Assay, Mouse Assay, Injection, Cross-linking Immunoprecipitation, Generated

    pLN-resident CD4 T cells exhibit an effector phenotype. 6–12-week-old C57BL/6 Foxp3-GFP mice were injected or not i.p. with 200 μg of anti-LFA-1 (αL) and anti-VLA-4 (α4) Abs. Forty-eight hours later, pLNs were harvested and analyzed. a CD62L, CD127, and CCR7 fluorescence histograms of CD4 Treg and CD4 Tmem cells from a representative treated and a representative control C57BL/6 Foxp3-GFP mouse. b Representative Ki-67 expression by CD4 Treg and CD4 Tmem cells from treated and control mice. Quantification is shown on the right side of the panel (unpaired t -test). Each dot represents an individual mouse. c Representative IL-10 expression by CD4 Treg cells and representative IL-2, IL-17, and IFN-γ expression by CD4 Tmem cells from treated and control mice. d Quantification of cytokine production by CD4 Treg and Tmem cells is shown as means ± SEM with paired t -test. The percentage of recovery for a given cytokine was calculated by dividing the absolute numbers of cells expressing this cytokine in treated mice by the mean absolute number of cells expressing this same cytokine in untreated animals (white bars). The gray bars correspond to the recovery of total CD4 Treg or CD4 Tmem cells in the same samples. * p

    Journal: Nature Communications

    Article Title: Profiling the lymphoid-resident T cell pool reveals modulation by age and microbiota

    doi: 10.1038/s41467-017-02458-4

    Figure Lengend Snippet: pLN-resident CD4 T cells exhibit an effector phenotype. 6–12-week-old C57BL/6 Foxp3-GFP mice were injected or not i.p. with 200 μg of anti-LFA-1 (αL) and anti-VLA-4 (α4) Abs. Forty-eight hours later, pLNs were harvested and analyzed. a CD62L, CD127, and CCR7 fluorescence histograms of CD4 Treg and CD4 Tmem cells from a representative treated and a representative control C57BL/6 Foxp3-GFP mouse. b Representative Ki-67 expression by CD4 Treg and CD4 Tmem cells from treated and control mice. Quantification is shown on the right side of the panel (unpaired t -test). Each dot represents an individual mouse. c Representative IL-10 expression by CD4 Treg cells and representative IL-2, IL-17, and IFN-γ expression by CD4 Tmem cells from treated and control mice. d Quantification of cytokine production by CD4 Treg and Tmem cells is shown as means ± SEM with paired t -test. The percentage of recovery for a given cytokine was calculated by dividing the absolute numbers of cells expressing this cytokine in treated mice by the mean absolute number of cells expressing this same cytokine in untreated animals (white bars). The gray bars correspond to the recovery of total CD4 Treg or CD4 Tmem cells in the same samples. * p

    Article Snippet: Blocking T cell entry into LNs and Pp C57BL/6 Foxp3-GFP mice were injected or not i.p. with 200 μg of anti-LFA-1 (clone M17/4) and anti-VLA-4 (clone PS/2) Abs (BioXcell) to block T cell entry into LNs and Peyer’s patches as previously described .

    Techniques: Mouse Assay, Injection, Fluorescence, Expressing

    T cell residence in SLOs increases with age in SPF mice. a Distribution of Treg, Tmem, and Tnaive cells among CD4 T cells in blood, pLNs (pooled superficial cervical, axillary, brachial, and inguinal LNs), mLNs, and spleen of 1–12-month-old C57BL/6 Foxp3-GFP mice. b C57BL/6 Foxp3-GFP mice of various ages were injected or not i.p. with 200 μg of anti-LFA-1 (αL) and anti-VLA-4 (α4) Abs. Forty-eight hours later, blood, pLNs, mLNs, and spleen were harvested and analyzed. Relative absolute numbers (recovery) of CD4 Treg, Tmem, and Tnaive cells in the indicated SLOs are shown as means ± SEM as a function of mouse age. c , d Percentages of Treg, Tmem, and Tnaive cells among total or “circulating” CD4 T cells are shown as means ± SEM for the indicated SLOs (see the experimental procedures for calculations)

    Journal: Nature Communications

    Article Title: Profiling the lymphoid-resident T cell pool reveals modulation by age and microbiota

    doi: 10.1038/s41467-017-02458-4

    Figure Lengend Snippet: T cell residence in SLOs increases with age in SPF mice. a Distribution of Treg, Tmem, and Tnaive cells among CD4 T cells in blood, pLNs (pooled superficial cervical, axillary, brachial, and inguinal LNs), mLNs, and spleen of 1–12-month-old C57BL/6 Foxp3-GFP mice. b C57BL/6 Foxp3-GFP mice of various ages were injected or not i.p. with 200 μg of anti-LFA-1 (αL) and anti-VLA-4 (α4) Abs. Forty-eight hours later, blood, pLNs, mLNs, and spleen were harvested and analyzed. Relative absolute numbers (recovery) of CD4 Treg, Tmem, and Tnaive cells in the indicated SLOs are shown as means ± SEM as a function of mouse age. c , d Percentages of Treg, Tmem, and Tnaive cells among total or “circulating” CD4 T cells are shown as means ± SEM for the indicated SLOs (see the experimental procedures for calculations)

    Article Snippet: Blocking T cell entry into LNs and Pp C57BL/6 Foxp3-GFP mice were injected or not i.p. with 200 μg of anti-LFA-1 (clone M17/4) and anti-VLA-4 (clone PS/2) Abs (BioXcell) to block T cell entry into LNs and Peyer’s patches as previously described .

    Techniques: Mouse Assay, Injection

    TGF-β suppression of EOMES-driven responses was common to CD4 T cells from mice infected with MCMV and HIV-infected patients. ( A and B ) Mixed chimeras with 1:1 ratio of WT (CD45.1, black) and ERCre + Tgfbr2 flox (CD45.2, red) bone marrow reconstituted prior to TGFβ-RII deletion, were infected with 2 × 10 4 PFU MCMV i.p. ( A ) Proportion of CD4 T cells expressing activation markers CD11a and CD49d over time after gating on congenic marker, is shown for postinfection day 14. ( B ) Overlays of EOMES and KLRG expression in activated CD4 T cells. ( C ) Upper panels show density plots of mRNA expression level vs. CD107 protein and quantification of Eomes and GrzB transcripts in CD107 + vs. CD107 – responding CD4 T cells. Lower panels show single-cell analysis of Eomes and granzyme B mRNA expression levels in HIV-specific IFN-γ + CD107a + CD4 T cells after a 5-hour Gag peptide stimulation using Biomark Fluidigm analysis. The transcript expression threshold (Et) was defined as the qPCR cycle number above background at which the transcript was detected. ( D and E ) Gag-responsive HIV-specific CD4 T cells were isolated and cultured for 5 days in the presence or absence of TGF-β and restimulated with GAG peptides in the presence of CD28/CD49d, and monensin for percentage of CD107 + CD4 T cells ( D ) and eomesodermin and granzyme B expression in HIV-specific CD107 + CD4 T cells ( E ). ( A and B ) Representative data from 2 independent experiments, with n = 4 or 5 mice/group. ( C – E ) Representative data from n = 10 HIV + treatment-naive subjects. Two-way ANOVA ( A ), paired t test ( B – E ), * P

    Journal: The Journal of Clinical Investigation

    Article Title: TGF- β receptor maintains CD4 T helper cell identity during chronic viral infections

    doi: 10.1172/JCI87041

    Figure Lengend Snippet: TGF-β suppression of EOMES-driven responses was common to CD4 T cells from mice infected with MCMV and HIV-infected patients. ( A and B ) Mixed chimeras with 1:1 ratio of WT (CD45.1, black) and ERCre + Tgfbr2 flox (CD45.2, red) bone marrow reconstituted prior to TGFβ-RII deletion, were infected with 2 × 10 4 PFU MCMV i.p. ( A ) Proportion of CD4 T cells expressing activation markers CD11a and CD49d over time after gating on congenic marker, is shown for postinfection day 14. ( B ) Overlays of EOMES and KLRG expression in activated CD4 T cells. ( C ) Upper panels show density plots of mRNA expression level vs. CD107 protein and quantification of Eomes and GrzB transcripts in CD107 + vs. CD107 – responding CD4 T cells. Lower panels show single-cell analysis of Eomes and granzyme B mRNA expression levels in HIV-specific IFN-γ + CD107a + CD4 T cells after a 5-hour Gag peptide stimulation using Biomark Fluidigm analysis. The transcript expression threshold (Et) was defined as the qPCR cycle number above background at which the transcript was detected. ( D and E ) Gag-responsive HIV-specific CD4 T cells were isolated and cultured for 5 days in the presence or absence of TGF-β and restimulated with GAG peptides in the presence of CD28/CD49d, and monensin for percentage of CD107 + CD4 T cells ( D ) and eomesodermin and granzyme B expression in HIV-specific CD107 + CD4 T cells ( E ). ( A and B ) Representative data from 2 independent experiments, with n = 4 or 5 mice/group. ( C – E ) Representative data from n = 10 HIV + treatment-naive subjects. Two-way ANOVA ( A ), paired t test ( B – E ), * P

    Article Snippet: Peripheral blood mononuclear cells (PBMCs) of HIV-infected individuals (chronically infected, treatment naive) were stimulated with HIV-Gag potential T cell epitope pools, anti-CD28, and anti-CD49d in the presence of anti–human CD107 and monensin (BD) for 5 hours.

    Techniques: Mouse Assay, Infection, Expressing, Activation Assay, Marker, Single-cell Analysis, Real-time Polymerase Chain Reaction, Isolation, Cell Culture

    Exclusive ablation of TGFβ-RII in CD4 T cells enhanced their numbers and terminal differentiation but limited LCMV-specific IgG1 during chronic infection. ( A ) Cd4-ERCre + Tgfbr2 fl/fl (iCD4-RII flox , red) mice, Cd4-ERCre + Tgfbr2 fl/+ heterozygotes (HET, gray) and Cre – littermate controls (WT, black) were infected with 2 × 10 6 PFU of LCMV Cl13. Blood was monitored by flow cytometry prior to infection ( A and B ) and at the indicated time points after infection ( C – G ). ( A ) TGFβ-RII expression over isotype (filled histogram) on B cells and CD4 and CD8 T cells prior to infection. Mean fluorescence intensity (MFI) for TGFβ-RII on CD4 T cells is graphed. ( B ) Percentage of activated CD44 + CD62L – CD4 T cells prior to infection. ( C ) Percentage and number of virus-specific PD1 + CD49d + cells over time after infection. ( D ) Percentage and number of PSGL1 + Ly6C + cells within activated CD4 T cells from C . ( E ) Percentage and number of EOMES- and granzyme B–expressing cells within activated CD4 T cells from C . ( F ) Expression overlay of indicated marker in EOMES + (black line) vs. EOMES – (gray fill) activated TGFβ-RII–deficient CD4 T cells from C . ( G ) Anti-LCMV (αLCMV) Ig levels in serum at postinfection day 30. ( H ) Viremia by plaque assay, as pooled results from 2 experiments. Representative of 3 independent experiments, with n = 4 or 5 mice/group. Two-way ANOVA ( A – H ), paired t test ( F ), * P

    Journal: The Journal of Clinical Investigation

    Article Title: TGF- β receptor maintains CD4 T helper cell identity during chronic viral infections

    doi: 10.1172/JCI87041

    Figure Lengend Snippet: Exclusive ablation of TGFβ-RII in CD4 T cells enhanced their numbers and terminal differentiation but limited LCMV-specific IgG1 during chronic infection. ( A ) Cd4-ERCre + Tgfbr2 fl/fl (iCD4-RII flox , red) mice, Cd4-ERCre + Tgfbr2 fl/+ heterozygotes (HET, gray) and Cre – littermate controls (WT, black) were infected with 2 × 10 6 PFU of LCMV Cl13. Blood was monitored by flow cytometry prior to infection ( A and B ) and at the indicated time points after infection ( C – G ). ( A ) TGFβ-RII expression over isotype (filled histogram) on B cells and CD4 and CD8 T cells prior to infection. Mean fluorescence intensity (MFI) for TGFβ-RII on CD4 T cells is graphed. ( B ) Percentage of activated CD44 + CD62L – CD4 T cells prior to infection. ( C ) Percentage and number of virus-specific PD1 + CD49d + cells over time after infection. ( D ) Percentage and number of PSGL1 + Ly6C + cells within activated CD4 T cells from C . ( E ) Percentage and number of EOMES- and granzyme B–expressing cells within activated CD4 T cells from C . ( F ) Expression overlay of indicated marker in EOMES + (black line) vs. EOMES – (gray fill) activated TGFβ-RII–deficient CD4 T cells from C . ( G ) Anti-LCMV (αLCMV) Ig levels in serum at postinfection day 30. ( H ) Viremia by plaque assay, as pooled results from 2 experiments. Representative of 3 independent experiments, with n = 4 or 5 mice/group. Two-way ANOVA ( A – H ), paired t test ( F ), * P

    Article Snippet: Peripheral blood mononuclear cells (PBMCs) of HIV-infected individuals (chronically infected, treatment naive) were stimulated with HIV-Gag potential T cell epitope pools, anti-CD28, and anti-CD49d in the presence of anti–human CD107 and monensin (BD) for 5 hours.

    Techniques: Infection, Mouse Assay, Flow Cytometry, Cytometry, Expressing, Fluorescence, Marker, Plaque Assay

    Expression of CXCR4 or CD49d (VLA-4) by different B-cell lines or normal blood B cells, as indicated. Displayed are fluorescence histograms depicting the relative red fluorescence intensity of the cells stained with anti-CXCR4 mAb’s ( a , shaded histograms) or with anti-CD49d mAb’s ( b , shaded histograms) compared with that of the same cells stained with a PE-conjugated isotype control mAb of irrelevant specificity (open histograms). The mean fluorescence intensity ratio of each specifically-stained cell population is displayed above the histograms.

    Journal: Journal of Clinical Investigation

    Article Title: Fibroblast-like synoviocytes support B-cell pseudoemperipolesis via a stromal cell-derived factor-1- and CD106 (VCAM-1)-dependent mechanism

    doi:

    Figure Lengend Snippet: Expression of CXCR4 or CD49d (VLA-4) by different B-cell lines or normal blood B cells, as indicated. Displayed are fluorescence histograms depicting the relative red fluorescence intensity of the cells stained with anti-CXCR4 mAb’s ( a , shaded histograms) or with anti-CD49d mAb’s ( b , shaded histograms) compared with that of the same cells stained with a PE-conjugated isotype control mAb of irrelevant specificity (open histograms). The mean fluorescence intensity ratio of each specifically-stained cell population is displayed above the histograms.

    Article Snippet: The following mAb’s specific for human surface antigens were used: anti-CXCR4 (12G5), anti-VCAM-1, anti-CD19, anti-CD20, anti-CD49d, and the appropriate isotype controls from PharMingen (San Diego, California, USA).

    Techniques: Expressing, Fluorescence, Staining

    The percentages of monocytes positive in FACS analysis for ‘fibronectin receptors’ (FN-FITC, see text), CD49e, CD49d, CD29, CD49b and CD49a integrins. Mean ± s.d. are given of original monocytes (first two columns of a set; □ healthy controls, ▪ patients) and of FN-adhered monocytes (last two columns of a set; healthy controls, patients) of healthy controls and patients. a = P

    Journal: Clinical and Experimental Immunology

    Article Title: An abnormal adherence of monocytes to fibronectin in thyroid autoimmunity has consequences for cell polarization and the development of veiled cells

    doi: 10.1046/j.1365-2249.2001.01583.x

    Figure Lengend Snippet: The percentages of monocytes positive in FACS analysis for ‘fibronectin receptors’ (FN-FITC, see text), CD49e, CD49d, CD29, CD49b and CD49a integrins. Mean ± s.d. are given of original monocytes (first two columns of a set; □ healthy controls, ▪ patients) and of FN-adhered monocytes (last two columns of a set; healthy controls, patients) of healthy controls and patients. a = P

    Article Snippet: The various monocyte fractions were analysed using a FACScan flow cytometer (Becton Dickinson) for the following markers using the following monoclonal antibodies: CD49a (VLA1, Serotec, Oxford, England); CD49b (VLA2, Serotec); CD49d (VLA4, Serotec); CD49e (VLA5, Serotec); CD29 (Beckman Coulter, Hialeah, FL, USA) and FITC-labelled FN.

    Techniques: FACS

    Characterization of MSCs derived from rat bone marrow and transfection efficiency of MSCs with pEF1/His C::IGF-1::EGFP. (A) Expression levels of surface markers in the isolated MSCs were measured by flow cytometry for CD29-FITC, CD54-PE, CD90-FITC, CD45-PE and CD49d-FITC. The red histograms represent staining with respective surface marker antibodies and the black histograms represent staining with isotype controls. (B) Prior to FACS, the transfection efficiency of MSCs with pEF1/His C::IGF-1::EGFP and the nonviral carrier PAM-ABP was ~20% (P4). (C) Following FACS, transfected MSCs were collected and the purity was increased to 95.1%. Therefore, approximately all MSCs expressed a green fluorescent signal and IGF-1. CD, cluster of differentiation; EGFP, enhanced green fluorescence protein; IGF, insulin-like growth factor; MSC, mesenchymal stem cells; FACS, fluorescence-activated cell sorting cell sorting; FITC, fluorescein isothiocyanate; PE, phycoerythrin.

    Journal: Molecular Medicine Reports

    Article Title: Therapeutic effects of a mesenchymal stem cell-based insulin-like growth factor-1/enhanced green fluorescent protein dual gene sorting system in a myocardial infarction rat model

    doi: 10.3892/mmr.2018.9561

    Figure Lengend Snippet: Characterization of MSCs derived from rat bone marrow and transfection efficiency of MSCs with pEF1/His C::IGF-1::EGFP. (A) Expression levels of surface markers in the isolated MSCs were measured by flow cytometry for CD29-FITC, CD54-PE, CD90-FITC, CD45-PE and CD49d-FITC. The red histograms represent staining with respective surface marker antibodies and the black histograms represent staining with isotype controls. (B) Prior to FACS, the transfection efficiency of MSCs with pEF1/His C::IGF-1::EGFP and the nonviral carrier PAM-ABP was ~20% (P4). (C) Following FACS, transfected MSCs were collected and the purity was increased to 95.1%. Therefore, approximately all MSCs expressed a green fluorescent signal and IGF-1. CD, cluster of differentiation; EGFP, enhanced green fluorescence protein; IGF, insulin-like growth factor; MSC, mesenchymal stem cells; FACS, fluorescence-activated cell sorting cell sorting; FITC, fluorescein isothiocyanate; PE, phycoerythrin.

    Article Snippet: Resuspended single cells were labeled with antibodies against surface markers, including anti-cluster of differentiation (CD)29-fluorescein isothiocyanate (FITC; cat. no. 102205; 1:50), anti-CD54-phycoerythrin (PE; cat. no. 202405; 1:80), anti-CD90-FITC (cat. no. 202504; 1:200), anti-CD45-PE (cat. no. 202207; 1:80), and anti-CD49d-FITC (cat. no. 200103; 1:200) antibodies (BioLegend, Inc., San Diego, CA, USA) in ice for 1 h. MSCs labeled with antibodies were sorted by flow cytometry.

    Techniques: Derivative Assay, Transfection, Expressing, Isolation, Flow Cytometry, Cytometry, Staining, Marker, FACS, Fluorescence

    Cytofluorimetric characterization of KMSC (A) and KMSC-derived MPs (B). One µm and 6 µm beads were used as internal size standards. P1 area was compatible with 1 µm. Red lines indicate the expression of annexin V, CD44, CD73, CD29, α4 integrin, α5 integrin, and α6 integrin surface molecules. Black lines indicate isotypic controls.

    Journal: PLoS ONE

    Article Title: Microparticles from Kidney-Derived Mesenchymal Stem Cells Act as Carriers of Proangiogenic Signals and Contribute to Recovery from Acute Kidney Injury

    doi: 10.1371/journal.pone.0087853

    Figure Lengend Snippet: Cytofluorimetric characterization of KMSC (A) and KMSC-derived MPs (B). One µm and 6 µm beads were used as internal size standards. P1 area was compatible with 1 µm. Red lines indicate the expression of annexin V, CD44, CD73, CD29, α4 integrin, α5 integrin, and α6 integrin surface molecules. Black lines indicate isotypic controls.

    Article Snippet: MPs were pre-incubated (15 min at 4°C) with blocking antibodies (1 µg/ml) against the identified adhesion molecules such as anti-α4 integrin, anti-α5 integrin, anti-α6 integrin, anti-CD29, CD44, or trypsin (0.5mM, Invitrogen) before incubation with the cells.

    Techniques: Derivative Assay, Expressing

    Incorporation of MPs in HUVEC. (A) Immunofluorescence analysis of HUVEC internalization (30 min at 37°C) of MPs labeled with PKH26 preincubated with or without trypsin (0.5 mM), or with 1 µg/ml blocking monoclonal antibody against CD44, CD29, α4 integrin, α5 integrin, and α6 integrin. (B) Representative FACS analyses of HUVEC internalization of MPs labeled with PKH26 (black curves) after 30 min of incubation at 37°C. MPs were preincubated with or without trypsin, or with 1 µg/ml blocking monoclonal antibodies against CD44, CD29, α4 integrin, α5 integrin, and α6 integrin. Black curves indicate internalization of untreated MPs. In the first panel, red curve indicates negative control (cells not incubated with MPs) and black dot curve indicates PKH 26 expression in cultured HUVEC treated with PKH 26 labeled MPs alone. In other panels, red curves indicate internalization of MPs in cultured HUVEC after pretreatment with trypsin or blocking antibodies. Δ denotes the degree of PKH 26 reduction after treatment with trypsin or blocking antibodies compared to PKH 26 labeled MPs alone. Three independent experiments were performed with similar results.

    Journal: PLoS ONE

    Article Title: Microparticles from Kidney-Derived Mesenchymal Stem Cells Act as Carriers of Proangiogenic Signals and Contribute to Recovery from Acute Kidney Injury

    doi: 10.1371/journal.pone.0087853

    Figure Lengend Snippet: Incorporation of MPs in HUVEC. (A) Immunofluorescence analysis of HUVEC internalization (30 min at 37°C) of MPs labeled with PKH26 preincubated with or without trypsin (0.5 mM), or with 1 µg/ml blocking monoclonal antibody against CD44, CD29, α4 integrin, α5 integrin, and α6 integrin. (B) Representative FACS analyses of HUVEC internalization of MPs labeled with PKH26 (black curves) after 30 min of incubation at 37°C. MPs were preincubated with or without trypsin, or with 1 µg/ml blocking monoclonal antibodies against CD44, CD29, α4 integrin, α5 integrin, and α6 integrin. Black curves indicate internalization of untreated MPs. In the first panel, red curve indicates negative control (cells not incubated with MPs) and black dot curve indicates PKH 26 expression in cultured HUVEC treated with PKH 26 labeled MPs alone. In other panels, red curves indicate internalization of MPs in cultured HUVEC after pretreatment with trypsin or blocking antibodies. Δ denotes the degree of PKH 26 reduction after treatment with trypsin or blocking antibodies compared to PKH 26 labeled MPs alone. Three independent experiments were performed with similar results.

    Article Snippet: MPs were pre-incubated (15 min at 4°C) with blocking antibodies (1 µg/ml) against the identified adhesion molecules such as anti-α4 integrin, anti-α5 integrin, anti-α6 integrin, anti-CD29, CD44, or trypsin (0.5mM, Invitrogen) before incubation with the cells.

    Techniques: Immunofluorescence, Labeling, Blocking Assay, FACS, Incubation, Negative Control, Expressing, Cell Culture