Structured Review

SLIT2 LTD pbmcs
Exogenous <t>Slit2</t> inhibits HIV-1 replication in T cells by binding to Robo1 receptor (a) Slit2 expression in partially purified conditioned media from Slit2-myc-transfected human embryonic kidney (HEK) cells (Slit2) or vector-transfected cells (VCs) was determined by WB analysis (left panel). HIV-1 IIIB replication in MT4 cells (middle panel) or PHA-stimulated peripheral blood mononuclear cells <t>(PBMCs)</t> from normal donors (right panel) in the presence of partially purified conditioned medium containing Slit2 (approximately 6 nmol/l) or conditioned medium from control HEK293 cells was determined by analyzing HIV-1 p24 antigen levels in the supernatants by ELISA. In the case of MT4 cells, the supernatants were collected after 48 h. Virus production in the positive control (HIV-1-infected MT4 cells cultured only with medium): 12.1 ng/ml p24 Ag. * P
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92/100 stars

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1) Product Images from "A novel role for Slit2/Robo1 axis in modulating HIV-1 replication in T cells"

Article Title: A novel role for Slit2/Robo1 axis in modulating HIV-1 replication in T cells

Journal: AIDS (London, England)

doi: 10.1097/QAD.0b013e32834bab86

Exogenous Slit2 inhibits HIV-1 replication in T cells by binding to Robo1 receptor (a) Slit2 expression in partially purified conditioned media from Slit2-myc-transfected human embryonic kidney (HEK) cells (Slit2) or vector-transfected cells (VCs) was determined by WB analysis (left panel). HIV-1 IIIB replication in MT4 cells (middle panel) or PHA-stimulated peripheral blood mononuclear cells (PBMCs) from normal donors (right panel) in the presence of partially purified conditioned medium containing Slit2 (approximately 6 nmol/l) or conditioned medium from control HEK293 cells was determined by analyzing HIV-1 p24 antigen levels in the supernatants by ELISA. In the case of MT4 cells, the supernatants were collected after 48 h. Virus production in the positive control (HIV-1-infected MT4 cells cultured only with medium): 12.1 ng/ml p24 Ag. * P
Figure Legend Snippet: Exogenous Slit2 inhibits HIV-1 replication in T cells by binding to Robo1 receptor (a) Slit2 expression in partially purified conditioned media from Slit2-myc-transfected human embryonic kidney (HEK) cells (Slit2) or vector-transfected cells (VCs) was determined by WB analysis (left panel). HIV-1 IIIB replication in MT4 cells (middle panel) or PHA-stimulated peripheral blood mononuclear cells (PBMCs) from normal donors (right panel) in the presence of partially purified conditioned medium containing Slit2 (approximately 6 nmol/l) or conditioned medium from control HEK293 cells was determined by analyzing HIV-1 p24 antigen levels in the supernatants by ELISA. In the case of MT4 cells, the supernatants were collected after 48 h. Virus production in the positive control (HIV-1-infected MT4 cells cultured only with medium): 12.1 ng/ml p24 Ag. * P

Techniques Used: Binding Assay, Expressing, Purification, Transfection, Plasmid Preparation, Western Blot, Enzyme-linked Immunosorbent Assay, Positive Control, Infection, Cell Culture

Overexpression and downregulation of Slit2 modulates HIV-1 replication (a) Slit2 expression in lysates from MT4 cells and Jurkat T cells loaded at different concentrations (left panel) and from peripheral blood mononuclear cells (PBMCs) isolated from three normal donors (right panel) was determined by western blot (WB) analysis. Equal protein loading was determined by probing the same blot with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody. (b) Slit2 expression in Jurkat T cells transfected with nontargeting (NT) or Slit2-specific small inhibitor RNA (siRNA) was analyzed by WB analysis (left panel). Intracellular p24 staining of the transfected cells infected with HIV-1 IIIB for 48 h was determined by flow cytometric analysis (right panel). * P
Figure Legend Snippet: Overexpression and downregulation of Slit2 modulates HIV-1 replication (a) Slit2 expression in lysates from MT4 cells and Jurkat T cells loaded at different concentrations (left panel) and from peripheral blood mononuclear cells (PBMCs) isolated from three normal donors (right panel) was determined by western blot (WB) analysis. Equal protein loading was determined by probing the same blot with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody. (b) Slit2 expression in Jurkat T cells transfected with nontargeting (NT) or Slit2-specific small inhibitor RNA (siRNA) was analyzed by WB analysis (left panel). Intracellular p24 staining of the transfected cells infected with HIV-1 IIIB for 48 h was determined by flow cytometric analysis (right panel). * P

Techniques Used: Over Expression, Expressing, Isolation, Western Blot, Transfection, Staining, Infection, Flow Cytometry

2) Product Images from "A novel role for Slit2/Robo1 axis in modulating HIV-1 replication in T cells"

Article Title: A novel role for Slit2/Robo1 axis in modulating HIV-1 replication in T cells

Journal: AIDS (London, England)

doi: 10.1097/QAD.0b013e32834bab86

Exogenous Slit2 inhibits HIV-1 replication in T cells by binding to Robo1 receptor (a) Slit2 expression in partially purified conditioned media from Slit2-myc-transfected human embryonic kidney (HEK) cells (Slit2) or vector-transfected cells (VCs) was determined by WB analysis (left panel). HIV-1 IIIB replication in MT4 cells (middle panel) or PHA-stimulated peripheral blood mononuclear cells (PBMCs) from normal donors (right panel) in the presence of partially purified conditioned medium containing Slit2 (approximately 6 nmol/l) or conditioned medium from control HEK293 cells was determined by analyzing HIV-1 p24 antigen levels in the supernatants by ELISA. In the case of MT4 cells, the supernatants were collected after 48 h. Virus production in the positive control (HIV-1-infected MT4 cells cultured only with medium): 12.1 ng/ml p24 Ag. * P
Figure Legend Snippet: Exogenous Slit2 inhibits HIV-1 replication in T cells by binding to Robo1 receptor (a) Slit2 expression in partially purified conditioned media from Slit2-myc-transfected human embryonic kidney (HEK) cells (Slit2) or vector-transfected cells (VCs) was determined by WB analysis (left panel). HIV-1 IIIB replication in MT4 cells (middle panel) or PHA-stimulated peripheral blood mononuclear cells (PBMCs) from normal donors (right panel) in the presence of partially purified conditioned medium containing Slit2 (approximately 6 nmol/l) or conditioned medium from control HEK293 cells was determined by analyzing HIV-1 p24 antigen levels in the supernatants by ELISA. In the case of MT4 cells, the supernatants were collected after 48 h. Virus production in the positive control (HIV-1-infected MT4 cells cultured only with medium): 12.1 ng/ml p24 Ag. * P

Techniques Used: Binding Assay, Expressing, Purification, Transfection, Plasmid Preparation, Western Blot, Enzyme-linked Immunosorbent Assay, Positive Control, Infection, Cell Culture

Overexpression and downregulation of Slit2 modulates HIV-1 replication (a) Slit2 expression in lysates from MT4 cells and Jurkat T cells loaded at different concentrations (left panel) and from peripheral blood mononuclear cells (PBMCs) isolated from three normal donors (right panel) was determined by western blot (WB) analysis. Equal protein loading was determined by probing the same blot with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody. (b) Slit2 expression in Jurkat T cells transfected with nontargeting (NT) or Slit2-specific small inhibitor RNA (siRNA) was analyzed by WB analysis (left panel). Intracellular p24 staining of the transfected cells infected with HIV-1 IIIB for 48 h was determined by flow cytometric analysis (right panel). * P
Figure Legend Snippet: Overexpression and downregulation of Slit2 modulates HIV-1 replication (a) Slit2 expression in lysates from MT4 cells and Jurkat T cells loaded at different concentrations (left panel) and from peripheral blood mononuclear cells (PBMCs) isolated from three normal donors (right panel) was determined by western blot (WB) analysis. Equal protein loading was determined by probing the same blot with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody. (b) Slit2 expression in Jurkat T cells transfected with nontargeting (NT) or Slit2-specific small inhibitor RNA (siRNA) was analyzed by WB analysis (left panel). Intracellular p24 staining of the transfected cells infected with HIV-1 IIIB for 48 h was determined by flow cytometric analysis (right panel). * P

Techniques Used: Over Expression, Expressing, Isolation, Western Blot, Transfection, Staining, Infection, Flow Cytometry

3) Product Images from "Fibroblasts secrete Slit2 to inhibit fibrocyte differentiation and fibrosis"

Article Title: Fibroblasts secrete Slit2 to inhibit fibrocyte differentiation and fibrosis

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.1417426112

Fractionation of FCM and identification of Slit2. ( A ) FCM was fractionated by centrifugation using 100-kDa filters. PBMCs were cultured for 5 d in the presence or absence of 10% (vol/vol) FCM, 10-fold concentrated 100-kDa retentate (Ret), or 100-kDa flow through (FT). After a 5-d incubation, cells were air-dried, fixed, and stained, and the number of fibrocytes was counted. Compared with PBMCs cultured in the absence of conditioned medium (SFM), FCM and the 100-kDa retentate significantly decreased the number of fibrocytes. Values are mean ± SEM ( n = 4). * P
Figure Legend Snippet: Fractionation of FCM and identification of Slit2. ( A ) FCM was fractionated by centrifugation using 100-kDa filters. PBMCs were cultured for 5 d in the presence or absence of 10% (vol/vol) FCM, 10-fold concentrated 100-kDa retentate (Ret), or 100-kDa flow through (FT). After a 5-d incubation, cells were air-dried, fixed, and stained, and the number of fibrocytes was counted. Compared with PBMCs cultured in the absence of conditioned medium (SFM), FCM and the 100-kDa retentate significantly decreased the number of fibrocytes. Values are mean ± SEM ( n = 4). * P

Techniques Used: Fractionation, Centrifugation, Cell Culture, Flow Cytometry, Incubation, Staining

The effect of recombinant Slit2 (rSlit2) on human fibrocyte differentiation. ( A ) Human PBMCs and isolated monocytes were cultured in the presence or absence of rSlit2 for 5 d. Cells were then air-dried, fixed, and stained, and fibrocytes were counted. Values are mean ± SEM ( n = 3). The absence of error bars indicates that the error was smaller than the plot symbol. rSlit2 concentrations at 0.1 ng/mL and above significantly inhibited fibrocyte differentiation ( t test). Lines are fits to sigmoidal dose–response curves with variable Hill coefficients. ( B ) MRC-5 FCM was incubated with beads labeled with anti-Slit2 antibodies, anti-HSP-47 (a control protein depletion), rabbit IgG (a control IgG), or unlabeled beads. Human PBMCs were cultured in the presence or absence of 10% (vol/vol) antibody-depleted FCM for 5 d. Values are mean ± SEM ( n = 4). * P
Figure Legend Snippet: The effect of recombinant Slit2 (rSlit2) on human fibrocyte differentiation. ( A ) Human PBMCs and isolated monocytes were cultured in the presence or absence of rSlit2 for 5 d. Cells were then air-dried, fixed, and stained, and fibrocytes were counted. Values are mean ± SEM ( n = 3). The absence of error bars indicates that the error was smaller than the plot symbol. rSlit2 concentrations at 0.1 ng/mL and above significantly inhibited fibrocyte differentiation ( t test). Lines are fits to sigmoidal dose–response curves with variable Hill coefficients. ( B ) MRC-5 FCM was incubated with beads labeled with anti-Slit2 antibodies, anti-HSP-47 (a control protein depletion), rabbit IgG (a control IgG), or unlabeled beads. Human PBMCs were cultured in the presence or absence of 10% (vol/vol) antibody-depleted FCM for 5 d. Values are mean ± SEM ( n = 4). * P

Techniques Used: Recombinant, Isolation, Cell Culture, Staining, Incubation, Labeling

4) Product Images from "A novel role for Slit2/Robo1 axis in modulating HIV-1 replication in T cells"

Article Title: A novel role for Slit2/Robo1 axis in modulating HIV-1 replication in T cells

Journal: AIDS (London, England)

doi: 10.1097/QAD.0b013e32834bab86

Exogenous Slit2 inhibits HIV-1 replication in T cells by binding to Robo1 receptor (a) Slit2 expression in partially purified conditioned media from Slit2-myc-transfected human embryonic kidney (HEK) cells (Slit2) or vector-transfected cells (VCs) was determined by WB analysis (left panel). HIV-1 IIIB replication in MT4 cells (middle panel) or PHA-stimulated peripheral blood mononuclear cells (PBMCs) from normal donors (right panel) in the presence of partially purified conditioned medium containing Slit2 (approximately 6 nmol/l) or conditioned medium from control HEK293 cells was determined by analyzing HIV-1 p24 antigen levels in the supernatants by ELISA. In the case of MT4 cells, the supernatants were collected after 48 h. Virus production in the positive control (HIV-1-infected MT4 cells cultured only with medium): 12.1 ng/ml p24 Ag. * P
Figure Legend Snippet: Exogenous Slit2 inhibits HIV-1 replication in T cells by binding to Robo1 receptor (a) Slit2 expression in partially purified conditioned media from Slit2-myc-transfected human embryonic kidney (HEK) cells (Slit2) or vector-transfected cells (VCs) was determined by WB analysis (left panel). HIV-1 IIIB replication in MT4 cells (middle panel) or PHA-stimulated peripheral blood mononuclear cells (PBMCs) from normal donors (right panel) in the presence of partially purified conditioned medium containing Slit2 (approximately 6 nmol/l) or conditioned medium from control HEK293 cells was determined by analyzing HIV-1 p24 antigen levels in the supernatants by ELISA. In the case of MT4 cells, the supernatants were collected after 48 h. Virus production in the positive control (HIV-1-infected MT4 cells cultured only with medium): 12.1 ng/ml p24 Ag. * P

Techniques Used: Binding Assay, Expressing, Purification, Transfection, Plasmid Preparation, Western Blot, Enzyme-linked Immunosorbent Assay, Positive Control, Infection, Cell Culture

Overexpression and downregulation of Slit2 modulates HIV-1 replication (a) Slit2 expression in lysates from MT4 cells and Jurkat T cells loaded at different concentrations (left panel) and from peripheral blood mononuclear cells (PBMCs) isolated from three normal donors (right panel) was determined by western blot (WB) analysis. Equal protein loading was determined by probing the same blot with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody. (b) Slit2 expression in Jurkat T cells transfected with nontargeting (NT) or Slit2-specific small inhibitor RNA (siRNA) was analyzed by WB analysis (left panel). Intracellular p24 staining of the transfected cells infected with HIV-1 IIIB for 48 h was determined by flow cytometric analysis (right panel). * P
Figure Legend Snippet: Overexpression and downregulation of Slit2 modulates HIV-1 replication (a) Slit2 expression in lysates from MT4 cells and Jurkat T cells loaded at different concentrations (left panel) and from peripheral blood mononuclear cells (PBMCs) isolated from three normal donors (right panel) was determined by western blot (WB) analysis. Equal protein loading was determined by probing the same blot with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody. (b) Slit2 expression in Jurkat T cells transfected with nontargeting (NT) or Slit2-specific small inhibitor RNA (siRNA) was analyzed by WB analysis (left panel). Intracellular p24 staining of the transfected cells infected with HIV-1 IIIB for 48 h was determined by flow cytometric analysis (right panel). * P

Techniques Used: Over Expression, Expressing, Isolation, Western Blot, Transfection, Staining, Infection, Flow Cytometry

Related Articles

Infection:

Article Title: A novel role for Slit2/Robo1 axis in modulating HIV-1 replication in T cells
Article Snippet: .. To further confirm the role of Slit2 in HIV infection, we overexpressed Slit2 in MT4 cells and PBMCs by transfecting pSecTagB-Slit2 construct [ ] or pSecTagB control vector in these cells. .. The efficiency of the transfection was confirmed by determining Slit2 expression in cell lysates by western blot analysis ( , left top panel).

Cell Culture:

Article Title: TNF-α–stimulated fibroblasts secrete lumican to promote fibrocyte differentiation
Article Snippet: .. To determine how lumican and Slit2 might compete to regulate fibrocyte differentiation, PBMC were cultured in SFM in the absence or presence of 10 μg/mL lumican and 500 pg/mL Slit2. .. Slit2 inhibited fibrocyte differentiation, lumican potentiated fibrocyte differentiation, and the addition of Slit2 was unable to block this effect of lumican on fibrocyte differentiation ( ).

Incubation:

Article Title: Potential Role of Axonal Chemorepellent Slit2 in Modulating Adventitial Inflammation in a Rat Carotid Artery Balloon Injury Model
Article Snippet: .. PBMCs were incubated for 3 hr in cell migration inserts (5 μm pore size) and 10 ng/ml of SDF-1 was added to the lower chambers in the presence and absence of CM from Ad/CMV/V5-, Ad/Slit2/V5- and Ad/Slit2-N/1118/V5-infected HEK 293 cells. .. SDF-1 induced a 3.4-fold increase in PBMC migration, and CM from HEK cells containing Slit2 and Slit2-N/1118 completely blocked SDF-1-induced PBMC migration, indicating that Slit2-N/1118 is a biologically active fragment ( ).

Inhibition:

Article Title: A novel role for Slit2/Robo1 axis in modulating HIV-1 replication in T cells
Article Snippet: .. Our studies indicate that Slit2 overexpression inhibited HIV-1 replication in MT4 and PBMCs, whereas inhibition of constitutive Slit2 expression in Jurkat T cells slightly enhanced HIV-1 replication, suggesting that endogenous Slit2 expression and secretion may modulate HIV-1 replication in T cells probably by acting in an autocrine/paracrine manner. .. We also showed that soluble Slit2 inhibited both X4-tropic and R5-tropic HIV-1 using both partially purified Slit2-conditioned medium and purified Slit2.

Construct:

Article Title: A novel role for Slit2/Robo1 axis in modulating HIV-1 replication in T cells
Article Snippet: .. To further confirm the role of Slit2 in HIV infection, we overexpressed Slit2 in MT4 cells and PBMCs by transfecting pSecTagB-Slit2 construct [ ] or pSecTagB control vector in these cells. .. The efficiency of the transfection was confirmed by determining Slit2 expression in cell lysates by western blot analysis ( , left top panel).

Expressing:

Article Title: A novel role for Slit2/Robo1 axis in modulating HIV-1 replication in T cells
Article Snippet: .. We also demonstrated Slit2 expression in PBMCs from normal donors ( , right panel). .. Next, we examined the effect of downregulation of endogenous Slit2 in Jurkat T cells using Slit2-specific siRNA on HIV-1 replication ( , left panel).

Article Title: A novel role for Slit2/Robo1 axis in modulating HIV-1 replication in T cells
Article Snippet: .. Our studies indicate that Slit2 overexpression inhibited HIV-1 replication in MT4 and PBMCs, whereas inhibition of constitutive Slit2 expression in Jurkat T cells slightly enhanced HIV-1 replication, suggesting that endogenous Slit2 expression and secretion may modulate HIV-1 replication in T cells probably by acting in an autocrine/paracrine manner. .. We also showed that soluble Slit2 inhibited both X4-tropic and R5-tropic HIV-1 using both partially purified Slit2-conditioned medium and purified Slit2.

Over Expression:

Article Title: A novel role for Slit2/Robo1 axis in modulating HIV-1 replication in T cells
Article Snippet: .. Our studies indicate that Slit2 overexpression inhibited HIV-1 replication in MT4 and PBMCs, whereas inhibition of constitutive Slit2 expression in Jurkat T cells slightly enhanced HIV-1 replication, suggesting that endogenous Slit2 expression and secretion may modulate HIV-1 replication in T cells probably by acting in an autocrine/paracrine manner. .. We also showed that soluble Slit2 inhibited both X4-tropic and R5-tropic HIV-1 using both partially purified Slit2-conditioned medium and purified Slit2.

Plasmid Preparation:

Article Title: A novel role for Slit2/Robo1 axis in modulating HIV-1 replication in T cells
Article Snippet: .. To further confirm the role of Slit2 in HIV infection, we overexpressed Slit2 in MT4 cells and PBMCs by transfecting pSecTagB-Slit2 construct [ ] or pSecTagB control vector in these cells. .. The efficiency of the transfection was confirmed by determining Slit2 expression in cell lysates by western blot analysis ( , left top panel).

Migration:

Article Title: Potential Role of Axonal Chemorepellent Slit2 in Modulating Adventitial Inflammation in a Rat Carotid Artery Balloon Injury Model
Article Snippet: .. PBMCs were incubated for 3 hr in cell migration inserts (5 μm pore size) and 10 ng/ml of SDF-1 was added to the lower chambers in the presence and absence of CM from Ad/CMV/V5-, Ad/Slit2/V5- and Ad/Slit2-N/1118/V5-infected HEK 293 cells. .. SDF-1 induced a 3.4-fold increase in PBMC migration, and CM from HEK cells containing Slit2 and Slit2-N/1118 completely blocked SDF-1-induced PBMC migration, indicating that Slit2-N/1118 is a biologically active fragment ( ).

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    SLIT2 LTD pbmcs
    Exogenous <t>Slit2</t> inhibits HIV-1 replication in T cells by binding to Robo1 receptor (a) Slit2 expression in partially purified conditioned media from Slit2-myc-transfected human embryonic kidney (HEK) cells (Slit2) or vector-transfected cells (VCs) was determined by WB analysis (left panel). HIV-1 IIIB replication in MT4 cells (middle panel) or PHA-stimulated peripheral blood mononuclear cells <t>(PBMCs)</t> from normal donors (right panel) in the presence of partially purified conditioned medium containing Slit2 (approximately 6 nmol/l) or conditioned medium from control HEK293 cells was determined by analyzing HIV-1 p24 antigen levels in the supernatants by ELISA. In the case of MT4 cells, the supernatants were collected after 48 h. Virus production in the positive control (HIV-1-infected MT4 cells cultured only with medium): 12.1 ng/ml p24 Ag. * P
    Pbmcs, supplied by SLIT2 LTD, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbmcs/product/SLIT2 LTD
    Average 92 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    pbmcs - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    85
    SLIT2 LTD slit2 transfected pbmcs
    Exogenous <t>Slit2</t> inhibits HIV-1 replication in T cells by binding to Robo1 receptor (a) Slit2 expression in partially purified conditioned media from <t>Slit2-myc-transfected</t> human embryonic kidney (HEK) cells (Slit2) or vector-transfected cells (VCs) was determined by WB analysis (left panel). HIV-1 IIIB replication in MT4 cells (middle panel) or PHA-stimulated peripheral blood mononuclear cells <t>(PBMCs)</t> from normal donors (right panel) in the presence of partially purified conditioned medium containing Slit2 (approximately 6 nmol/l) or conditioned medium from control HEK293 cells was determined by analyzing HIV-1 p24 antigen levels in the supernatants by ELISA. In the case of MT4 cells, the supernatants were collected after 48 h. Virus production in the positive control (HIV-1-infected MT4 cells cultured only with medium): 12.1 ng/ml p24 Ag. * P
    Slit2 Transfected Pbmcs, supplied by SLIT2 LTD, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/slit2 transfected pbmcs/product/SLIT2 LTD
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    slit2 transfected pbmcs - by Bioz Stars, 2020-09
    85/100 stars
      Buy from Supplier

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    Exogenous Slit2 inhibits HIV-1 replication in T cells by binding to Robo1 receptor (a) Slit2 expression in partially purified conditioned media from Slit2-myc-transfected human embryonic kidney (HEK) cells (Slit2) or vector-transfected cells (VCs) was determined by WB analysis (left panel). HIV-1 IIIB replication in MT4 cells (middle panel) or PHA-stimulated peripheral blood mononuclear cells (PBMCs) from normal donors (right panel) in the presence of partially purified conditioned medium containing Slit2 (approximately 6 nmol/l) or conditioned medium from control HEK293 cells was determined by analyzing HIV-1 p24 antigen levels in the supernatants by ELISA. In the case of MT4 cells, the supernatants were collected after 48 h. Virus production in the positive control (HIV-1-infected MT4 cells cultured only with medium): 12.1 ng/ml p24 Ag. * P

    Journal: AIDS (London, England)

    Article Title: A novel role for Slit2/Robo1 axis in modulating HIV-1 replication in T cells

    doi: 10.1097/QAD.0b013e32834bab86

    Figure Lengend Snippet: Exogenous Slit2 inhibits HIV-1 replication in T cells by binding to Robo1 receptor (a) Slit2 expression in partially purified conditioned media from Slit2-myc-transfected human embryonic kidney (HEK) cells (Slit2) or vector-transfected cells (VCs) was determined by WB analysis (left panel). HIV-1 IIIB replication in MT4 cells (middle panel) or PHA-stimulated peripheral blood mononuclear cells (PBMCs) from normal donors (right panel) in the presence of partially purified conditioned medium containing Slit2 (approximately 6 nmol/l) or conditioned medium from control HEK293 cells was determined by analyzing HIV-1 p24 antigen levels in the supernatants by ELISA. In the case of MT4 cells, the supernatants were collected after 48 h. Virus production in the positive control (HIV-1-infected MT4 cells cultured only with medium): 12.1 ng/ml p24 Ag. * P

    Article Snippet: Our studies indicate that Slit2 overexpression inhibited HIV-1 replication in MT4 and PBMCs, whereas inhibition of constitutive Slit2 expression in Jurkat T cells slightly enhanced HIV-1 replication, suggesting that endogenous Slit2 expression and secretion may modulate HIV-1 replication in T cells probably by acting in an autocrine/paracrine manner.

    Techniques: Binding Assay, Expressing, Purification, Transfection, Plasmid Preparation, Western Blot, Enzyme-linked Immunosorbent Assay, Positive Control, Infection, Cell Culture

    Overexpression and downregulation of Slit2 modulates HIV-1 replication (a) Slit2 expression in lysates from MT4 cells and Jurkat T cells loaded at different concentrations (left panel) and from peripheral blood mononuclear cells (PBMCs) isolated from three normal donors (right panel) was determined by western blot (WB) analysis. Equal protein loading was determined by probing the same blot with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody. (b) Slit2 expression in Jurkat T cells transfected with nontargeting (NT) or Slit2-specific small inhibitor RNA (siRNA) was analyzed by WB analysis (left panel). Intracellular p24 staining of the transfected cells infected with HIV-1 IIIB for 48 h was determined by flow cytometric analysis (right panel). * P

    Journal: AIDS (London, England)

    Article Title: A novel role for Slit2/Robo1 axis in modulating HIV-1 replication in T cells

    doi: 10.1097/QAD.0b013e32834bab86

    Figure Lengend Snippet: Overexpression and downregulation of Slit2 modulates HIV-1 replication (a) Slit2 expression in lysates from MT4 cells and Jurkat T cells loaded at different concentrations (left panel) and from peripheral blood mononuclear cells (PBMCs) isolated from three normal donors (right panel) was determined by western blot (WB) analysis. Equal protein loading was determined by probing the same blot with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody. (b) Slit2 expression in Jurkat T cells transfected with nontargeting (NT) or Slit2-specific small inhibitor RNA (siRNA) was analyzed by WB analysis (left panel). Intracellular p24 staining of the transfected cells infected with HIV-1 IIIB for 48 h was determined by flow cytometric analysis (right panel). * P

    Article Snippet: Our studies indicate that Slit2 overexpression inhibited HIV-1 replication in MT4 and PBMCs, whereas inhibition of constitutive Slit2 expression in Jurkat T cells slightly enhanced HIV-1 replication, suggesting that endogenous Slit2 expression and secretion may modulate HIV-1 replication in T cells probably by acting in an autocrine/paracrine manner.

    Techniques: Over Expression, Expressing, Isolation, Western Blot, Transfection, Staining, Infection, Flow Cytometry

    Fractionation of FCM+TNF-α and identification of lumican. ( A ) FCM+TNF-α was collected and analyzed for cytokines. ( B ) FCM+TNF-α was fractionated by centrifugation using 100-, 10-, and 3-kDa cutoff filters. PBMC were cultured for

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: TNF-α–stimulated fibroblasts secrete lumican to promote fibrocyte differentiation

    doi: 10.1073/pnas.1507387112

    Figure Lengend Snippet: Fractionation of FCM+TNF-α and identification of lumican. ( A ) FCM+TNF-α was collected and analyzed for cytokines. ( B ) FCM+TNF-α was fractionated by centrifugation using 100-, 10-, and 3-kDa cutoff filters. PBMC were cultured for

    Article Snippet: To determine how lumican and Slit2 might compete to regulate fibrocyte differentiation, PBMC were cultured in SFM in the absence or presence of 10 μg/mL lumican and 500 pg/mL Slit2.

    Techniques: Fractionation, Centrifugation, Cell Culture

    Lumican potentiates human fibrocyte differentiation. ( A ) Human PBMC and isolated monocytes were cultured in the presence or absence of recombinant lumican for 5 d. Cells were then air-dried, fixed, stained, and fibrocytes were counted. Values are mean

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: TNF-α–stimulated fibroblasts secrete lumican to promote fibrocyte differentiation

    doi: 10.1073/pnas.1507387112

    Figure Lengend Snippet: Lumican potentiates human fibrocyte differentiation. ( A ) Human PBMC and isolated monocytes were cultured in the presence or absence of recombinant lumican for 5 d. Cells were then air-dried, fixed, stained, and fibrocytes were counted. Values are mean

    Article Snippet: To determine how lumican and Slit2 might compete to regulate fibrocyte differentiation, PBMC were cultured in SFM in the absence or presence of 10 μg/mL lumican and 500 pg/mL Slit2.

    Techniques: Isolation, Cell Culture, Recombinant, Staining

    α2 integrin small molecule inhibitor BTT 3033 inhibits lumican-induced fibrocyte differentiation. PBMC were incubated in the presence or absence of 500 nM BTT3033 for 30 min, and then cultured in the presence or absence of 10 μg/mL lumican

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: TNF-α–stimulated fibroblasts secrete lumican to promote fibrocyte differentiation

    doi: 10.1073/pnas.1507387112

    Figure Lengend Snippet: α2 integrin small molecule inhibitor BTT 3033 inhibits lumican-induced fibrocyte differentiation. PBMC were incubated in the presence or absence of 500 nM BTT3033 for 30 min, and then cultured in the presence or absence of 10 μg/mL lumican

    Article Snippet: To determine how lumican and Slit2 might compete to regulate fibrocyte differentiation, PBMC were cultured in SFM in the absence or presence of 10 μg/mL lumican and 500 pg/mL Slit2.

    Techniques: Incubation, Cell Culture

    The effect of lumican on monocyte differentiation and macrophage polarization. ( A ) Human PBMC were cultured in the presence or absence of recombinant lumican for 6 d. ( B ) Human PBMC were cultured for 6 d, the medium was then replaced with medium in the

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: TNF-α–stimulated fibroblasts secrete lumican to promote fibrocyte differentiation

    doi: 10.1073/pnas.1507387112

    Figure Lengend Snippet: The effect of lumican on monocyte differentiation and macrophage polarization. ( A ) Human PBMC were cultured in the presence or absence of recombinant lumican for 6 d. ( B ) Human PBMC were cultured for 6 d, the medium was then replaced with medium in the

    Article Snippet: To determine how lumican and Slit2 might compete to regulate fibrocyte differentiation, PBMC were cultured in SFM in the absence or presence of 10 μg/mL lumican and 500 pg/mL Slit2.

    Techniques: Cell Culture, Recombinant

    The competition of lumican with SAP or Slit2. ( A ) PBMC were incubated with increasing concentrations of FCM+TNF-α in the presence of 0, 4. or 60 μg/mL SAP. Values are mean ± SEM ( n = 4). ( B ) PBMC were incubated with increasing

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: TNF-α–stimulated fibroblasts secrete lumican to promote fibrocyte differentiation

    doi: 10.1073/pnas.1507387112

    Figure Lengend Snippet: The competition of lumican with SAP or Slit2. ( A ) PBMC were incubated with increasing concentrations of FCM+TNF-α in the presence of 0, 4. or 60 μg/mL SAP. Values are mean ± SEM ( n = 4). ( B ) PBMC were incubated with increasing

    Article Snippet: To determine how lumican and Slit2 might compete to regulate fibrocyte differentiation, PBMC were cultured in SFM in the absence or presence of 10 μg/mL lumican and 500 pg/mL Slit2.

    Techniques: Incubation

    Some anti-integrin antibodies inhibit lumican-induced fibrocyte differentiation. PBMC were incubated with 5 μg/mL of mouse IgG1 or the indicated anti-integrin antibodies (specific integrin Ab in parenthesis), and then cultured in the presence

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: TNF-α–stimulated fibroblasts secrete lumican to promote fibrocyte differentiation

    doi: 10.1073/pnas.1507387112

    Figure Lengend Snippet: Some anti-integrin antibodies inhibit lumican-induced fibrocyte differentiation. PBMC were incubated with 5 μg/mL of mouse IgG1 or the indicated anti-integrin antibodies (specific integrin Ab in parenthesis), and then cultured in the presence

    Article Snippet: To determine how lumican and Slit2 might compete to regulate fibrocyte differentiation, PBMC were cultured in SFM in the absence or presence of 10 μg/mL lumican and 500 pg/mL Slit2.

    Techniques: Incubation, Cell Culture

    Exogenous Slit2 inhibits HIV-1 replication in T cells by binding to Robo1 receptor (a) Slit2 expression in partially purified conditioned media from Slit2-myc-transfected human embryonic kidney (HEK) cells (Slit2) or vector-transfected cells (VCs) was determined by WB analysis (left panel). HIV-1 IIIB replication in MT4 cells (middle panel) or PHA-stimulated peripheral blood mononuclear cells (PBMCs) from normal donors (right panel) in the presence of partially purified conditioned medium containing Slit2 (approximately 6 nmol/l) or conditioned medium from control HEK293 cells was determined by analyzing HIV-1 p24 antigen levels in the supernatants by ELISA. In the case of MT4 cells, the supernatants were collected after 48 h. Virus production in the positive control (HIV-1-infected MT4 cells cultured only with medium): 12.1 ng/ml p24 Ag. * P

    Journal: AIDS (London, England)

    Article Title: A novel role for Slit2/Robo1 axis in modulating HIV-1 replication in T cells

    doi: 10.1097/QAD.0b013e32834bab86

    Figure Lengend Snippet: Exogenous Slit2 inhibits HIV-1 replication in T cells by binding to Robo1 receptor (a) Slit2 expression in partially purified conditioned media from Slit2-myc-transfected human embryonic kidney (HEK) cells (Slit2) or vector-transfected cells (VCs) was determined by WB analysis (left panel). HIV-1 IIIB replication in MT4 cells (middle panel) or PHA-stimulated peripheral blood mononuclear cells (PBMCs) from normal donors (right panel) in the presence of partially purified conditioned medium containing Slit2 (approximately 6 nmol/l) or conditioned medium from control HEK293 cells was determined by analyzing HIV-1 p24 antigen levels in the supernatants by ELISA. In the case of MT4 cells, the supernatants were collected after 48 h. Virus production in the positive control (HIV-1-infected MT4 cells cultured only with medium): 12.1 ng/ml p24 Ag. * P

    Article Snippet: The Slit2-transfected MT4 cells showed approximately two-fold inhibition of HIV-1 replication ( , right panel), whereas the Slit2-transfected PBMCs showed an inhibition of approximately 3.5-fold using HIV-1 IIIB ( , left panel) and approximately two-fold using HIV-1 BaL ( , right panel) in comparison with vector-transfected cells, suggesting that overexpression of Slit2 in T cells was protective against HIV-1 infection.

    Techniques: Binding Assay, Expressing, Purification, Transfection, Plasmid Preparation, Western Blot, Enzyme-linked Immunosorbent Assay, Positive Control, Infection, Cell Culture

    Overexpression and downregulation of Slit2 modulates HIV-1 replication (a) Slit2 expression in lysates from MT4 cells and Jurkat T cells loaded at different concentrations (left panel) and from peripheral blood mononuclear cells (PBMCs) isolated from three normal donors (right panel) was determined by western blot (WB) analysis. Equal protein loading was determined by probing the same blot with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody. (b) Slit2 expression in Jurkat T cells transfected with nontargeting (NT) or Slit2-specific small inhibitor RNA (siRNA) was analyzed by WB analysis (left panel). Intracellular p24 staining of the transfected cells infected with HIV-1 IIIB for 48 h was determined by flow cytometric analysis (right panel). * P

    Journal: AIDS (London, England)

    Article Title: A novel role for Slit2/Robo1 axis in modulating HIV-1 replication in T cells

    doi: 10.1097/QAD.0b013e32834bab86

    Figure Lengend Snippet: Overexpression and downregulation of Slit2 modulates HIV-1 replication (a) Slit2 expression in lysates from MT4 cells and Jurkat T cells loaded at different concentrations (left panel) and from peripheral blood mononuclear cells (PBMCs) isolated from three normal donors (right panel) was determined by western blot (WB) analysis. Equal protein loading was determined by probing the same blot with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody. (b) Slit2 expression in Jurkat T cells transfected with nontargeting (NT) or Slit2-specific small inhibitor RNA (siRNA) was analyzed by WB analysis (left panel). Intracellular p24 staining of the transfected cells infected with HIV-1 IIIB for 48 h was determined by flow cytometric analysis (right panel). * P

    Article Snippet: The Slit2-transfected MT4 cells showed approximately two-fold inhibition of HIV-1 replication ( , right panel), whereas the Slit2-transfected PBMCs showed an inhibition of approximately 3.5-fold using HIV-1 IIIB ( , left panel) and approximately two-fold using HIV-1 BaL ( , right panel) in comparison with vector-transfected cells, suggesting that overexpression of Slit2 in T cells was protective against HIV-1 infection.

    Techniques: Over Expression, Expressing, Isolation, Western Blot, Transfection, Staining, Infection, Flow Cytometry