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Santa Cruz Biotechnology mouse integrin α4 shrna

Role of VLA-4 <t>integrin</t> in PD-L1 expression downregulation by SAS. ( A ) B16/F10 melanoma cells or ( B ) D122 cells were cultured on vascular cell adhesion molecule-1 (VCAM-1)- or bovine serum albumin (BSA)-coated plates with or without SAS for 1 h. The cells were washed twice . The percentage of attached cells, representing VLA-4 (very late antigen- <t>4</t> ) activity, was determined by the XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) viability test relative to the control PBS. *p<0.001 vs. BSA; #p<0.01 vs. PBS; **p<0.001 vs. PBS. Significance was calculated via one-way ANOVA. The results are presented as the mean+SE of 3 experiments. ( C ) Human VLA-4-negative AML cells isolated from AML patients were cultured on FN-coated plates for 24 hours. The cells were collected and stained with PE-conjugated anti-mouse PD-L1. ( D ) B16/F10 melanoma cells were transfected with either control or (E) VLA-4 <t>shRNA</t> (short hairpin RNA) and cultured with or without different concentrations of SAS. The cells were collected and stained with either PE-conjugated anti-mouse PD-L1 antibodies or isotype-matched controls. The results show one representative of 3 experiments. Figs C, D and E are accompanied by quantitative data from flow cytometry analysis. # p<0.001 vs. the isotype control; *p<0.05 vs. the PBS group. **p<0.01 vs. PBS.

Mouse Integrin α4 Shrna, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse integrin α4 shrna - by Bioz Stars, 2026-03

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1) Product Images from "Inhibition of α4β1 Integrin Activity by Small Tellurium Compounds Regulates PD-L1 Expression and Enhances Antitumor Effects"

Article Title: Inhibition of α4β1 Integrin Activity by Small Tellurium Compounds Regulates PD-L1 Expression and Enhances Antitumor Effects

Journal: International Journal of Biological Sciences

doi: 10.7150/ijbs.95350

Role of VLA-4 integrin in PD-L1 expression downregulation by SAS. ( A ) B16/F10 melanoma cells or ( B ) D122 cells were cultured on vascular cell adhesion molecule-1 (VCAM-1)- or bovine serum albumin (BSA)-coated plates with or without SAS for 1 h. The cells were washed twice . The percentage of attached cells, representing VLA-4 (very late antigen- 4 ) activity, was determined by the XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) viability test relative to the control PBS. *p<0.001 vs. BSA; #p<0.01 vs. PBS; **p<0.001 vs. PBS. Significance was calculated via one-way ANOVA. The results are presented as the mean+SE of 3 experiments. ( C ) Human VLA-4-negative AML cells isolated from AML patients were cultured on FN-coated plates for 24 hours. The cells were collected and stained with PE-conjugated anti-mouse PD-L1. ( D ) B16/F10 melanoma cells were transfected with either control or (E) VLA-4 shRNA (short hairpin RNA) and cultured with or without different concentrations of SAS. The cells were collected and stained with either PE-conjugated anti-mouse PD-L1 antibodies or isotype-matched controls. The results show one representative of 3 experiments. Figs C, D and E are accompanied by quantitative data from flow cytometry analysis. # p<0.001 vs. the isotype control; *p<0.05 vs. the PBS group. **p<0.01 vs. PBS.

Figure Legend Snippet: Role of VLA-4 integrin in PD-L1 expression downregulation by SAS. ( A ) B16/F10 melanoma cells or ( B ) D122 cells were cultured on vascular cell adhesion molecule-1 (VCAM-1)- or bovine serum albumin (BSA)-coated plates with or without SAS for 1 h. The cells were washed twice . The percentage of attached cells, representing VLA-4 (very late antigen- 4 ) activity, was determined by the XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) viability test relative to the control PBS. *p<0.001 vs. BSA; #p<0.01 vs. PBS; **p<0.001 vs. PBS. Significance was calculated via one-way ANOVA. The results are presented as the mean+SE of 3 experiments. ( C ) Human VLA-4-negative AML cells isolated from AML patients were cultured on FN-coated plates for 24 hours. The cells were collected and stained with PE-conjugated anti-mouse PD-L1. ( D ) B16/F10 melanoma cells were transfected with either control or (E) VLA-4 shRNA (short hairpin RNA) and cultured with or without different concentrations of SAS. The cells were collected and stained with either PE-conjugated anti-mouse PD-L1 antibodies or isotype-matched controls. The results show one representative of 3 experiments. Figs C, D and E are accompanied by quantitative data from flow cytometry analysis. # p<0.001 vs. the isotype control; *p<0.05 vs. the PBS group. **p<0.01 vs. PBS.

Techniques Used: Expressing, Cell Culture, Activity Assay, Control, Isolation, Staining, Transfection, shRNA, Flow Cytometry

SAS inhibits both PD-L1 expression and VLA-4 activity in U937 myelomonocytic human leukemic cells. ( A ) U937 human cells were cultured in the presence or absence of SAS. The cells were collected and stained with either PE-conjugated anti-human PD-L1 antibodies or their respective isotype-matched controls. The results show one representative experiment of 3 performed. ( B ) U937 cells were cultured on VCAM-1- or BSA-coated plates with or without SAS for 1 h. The cells were subsequently washed twice. The percentage of attached cells (representing VLA-4 activity) was determined by the XTT viability test relative to the control PBS. * p<0.001 vs. BSA **p<0.001 vs. PBS. Significance was calculated via one-way ANOVA. The results represent the mean +SE from 3 experiments ( B ). ( C ) The VLA-4 conformational structure was detected by FRET. VLA-4 activation involves the separation of the fluorescently tagged cytoplasmic ends of the α- and β-subunits of the integrin, resulting in a reduction in the FRET signal. U937 cells were transfected with α4-mCFP and β1-mYFP. After 48 hours, the cells were activated and fixed. FRET of the molecular interaction between the cytoplasmic domains of α4 and β1 was determined as described in the Materials and Methods. ( D ) Results for FRET positive controls (cells expressing CFP and YFP encoded on the same plasmid (i.e., maximal FRET obtained in this system) and negative controls (CFP and YFP expressed by different plasmids and undergoing minimal FRET, i.e., only that produced by random colocalizations). *p<0.01 vs. the negative control. Significance was calculated via a two-tailed t test. PLL (poly-L-lysine). FN (Fibronectin).

Figure Legend Snippet: SAS inhibits both PD-L1 expression and VLA-4 activity in U937 myelomonocytic human leukemic cells. ( A ) U937 human cells were cultured in the presence or absence of SAS. The cells were collected and stained with either PE-conjugated anti-human PD-L1 antibodies or their respective isotype-matched controls. The results show one representative experiment of 3 performed. ( B ) U937 cells were cultured on VCAM-1- or BSA-coated plates with or without SAS for 1 h. The cells were subsequently washed twice. The percentage of attached cells (representing VLA-4 activity) was determined by the XTT viability test relative to the control PBS. * p<0.001 vs. BSA **p<0.001 vs. PBS. Significance was calculated via one-way ANOVA. The results represent the mean +SE from 3 experiments ( B ). ( C ) The VLA-4 conformational structure was detected by FRET. VLA-4 activation involves the separation of the fluorescently tagged cytoplasmic ends of the α- and β-subunits of the integrin, resulting in a reduction in the FRET signal. U937 cells were transfected with α4-mCFP and β1-mYFP. After 48 hours, the cells were activated and fixed. FRET of the molecular interaction between the cytoplasmic domains of α4 and β1 was determined as described in the Materials and Methods. ( D ) Results for FRET positive controls (cells expressing CFP and YFP encoded on the same plasmid (i.e., maximal FRET obtained in this system) and negative controls (CFP and YFP expressed by different plasmids and undergoing minimal FRET, i.e., only that produced by random colocalizations). *p<0.01 vs. the negative control. Significance was calculated via a two-tailed t test. PLL (poly-L-lysine). FN (Fibronectin).

Techniques Used: Expressing, Activity Assay, Cell Culture, Staining, Control, Activation Assay, Transfection, Plasmid Preparation, Produced, Negative Control, Two Tailed Test

Associations between VLA-4, IL-10 and PD-L1. ( A ) B16/F10 cells were cultured on FN-coated plates in the presence or absence of SAS at various concentrations. The cells were collected, fixed, permeabilized and stained with PE-conjugated αIL-10 Abs. ( B ) SK-MEL23 cells were cultured on FN-coated plates in the presence or absence of SAS at various concentrations, with or without IFNγ (2 μg/ml), for 48 h. The supernatants were collected, and IL-10 levels were measured by ELISA. *p<0.0001 vs. without IFNγ; **p<0.001 vs. PBS. Significance was calculated via two-way ANOVA. The results represent the mean+SE of 3 experiments. SK-MEL23 cells without ( C, E ) or with IFNγ ( D, F ) were cultured on FN-coated plates in the presence or absence of SAS at various concentrations for 24 h, after which the cells were collected and stained for PD-L1 ( C, D ) or VLA-4 ( E, F ). All FACS results show one representative of 3 experiments. ( G ) SK-MEL23 cells were cultured in the presence or absence of SAS at various concentrations with or without IFNγ (2 μg/ml) for 24 h. The cells were detached and replated again on VCAM-1- or BSA-coated plates with or without SAS at various concentrations for 1 h. The cells were subsequently washed twice . The percentage of attached cells (representing VLA-4 activity) was determined by the XTT viability test relative to the control. *p<0.0001 vs. without IFNγ; **p<0.001 vs. PBS. Significance was calculated via two-way ANOVA. The results represent the mean+SE from 3 experiments. VLA-4 (Very late antigen-4/integrin α4β1). IFNγ (interferon gamma).

Figure Legend Snippet: Associations between VLA-4, IL-10 and PD-L1. ( A ) B16/F10 cells were cultured on FN-coated plates in the presence or absence of SAS at various concentrations. The cells were collected, fixed, permeabilized and stained with PE-conjugated αIL-10 Abs. ( B ) SK-MEL23 cells were cultured on FN-coated plates in the presence or absence of SAS at various concentrations, with or without IFNγ (2 μg/ml), for 48 h. The supernatants were collected, and IL-10 levels were measured by ELISA. *p<0.0001 vs. without IFNγ; **p<0.001 vs. PBS. Significance was calculated via two-way ANOVA. The results represent the mean+SE of 3 experiments. SK-MEL23 cells without ( C, E ) or with IFNγ ( D, F ) were cultured on FN-coated plates in the presence or absence of SAS at various concentrations for 24 h, after which the cells were collected and stained for PD-L1 ( C, D ) or VLA-4 ( E, F ). All FACS results show one representative of 3 experiments. ( G ) SK-MEL23 cells were cultured in the presence or absence of SAS at various concentrations with or without IFNγ (2 μg/ml) for 24 h. The cells were detached and replated again on VCAM-1- or BSA-coated plates with or without SAS at various concentrations for 1 h. The cells were subsequently washed twice . The percentage of attached cells (representing VLA-4 activity) was determined by the XTT viability test relative to the control. *p<0.0001 vs. without IFNγ; **p<0.001 vs. PBS. Significance was calculated via two-way ANOVA. The results represent the mean+SE from 3 experiments. VLA-4 (Very late antigen-4/integrin α4β1). IFNγ (interferon gamma).

Techniques Used: Cell Culture, Staining, Enzyme-linked Immunosorbent Assay, Activity Assay, Control

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Journal: International Journal of Biological Sciences

Article Title: Inhibition of α4β1 Integrin Activity by Small Tellurium Compounds Regulates PD-L1 Expression and Enhances Antitumor Effects

doi: 10.7150/ijbs.95350

Figure Lengend Snippet: Role of VLA-4 integrin in PD-L1 expression downregulation by SAS. ( A ) B16/F10 melanoma cells or ( B ) D122 cells were cultured on vascular cell adhesion molecule-1 (VCAM-1)- or bovine serum albumin (BSA)-coated plates with or without SAS for 1 h. The cells were washed twice . The percentage of attached cells, representing VLA-4 (very late antigen- 4 ) activity, was determined by the XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) viability test relative to the control PBS. *p<0.001 vs. BSA; #p<0.01 vs. PBS; **p<0.001 vs. PBS. Significance was calculated via one-way ANOVA. The results are presented as the mean+SE of 3 experiments. ( C ) Human VLA-4-negative AML cells isolated from AML patients were cultured on FN-coated plates for 24 hours. The cells were collected and stained with PE-conjugated anti-mouse PD-L1. ( D ) B16/F10 melanoma cells were transfected with either control or (E) VLA-4 shRNA (short hairpin RNA) and cultured with or without different concentrations of SAS. The cells were collected and stained with either PE-conjugated anti-mouse PD-L1 antibodies or isotype-matched controls. The results show one representative of 3 experiments. Figs C, D and E are accompanied by quantitative data from flow cytometry analysis. # p<0.001 vs. the isotype control; *p<0.05 vs. the PBS group. **p<0.01 vs. PBS.

Article Snippet: USA); paclitaxel (Sigma Aldrich); p65-GFP-RelA (Addgene; Watertown, MA, USA); p239-AKT (NIH); VLA-4 shRNA, mouse integrin α4 shRNA, and control plasmid (Santa Cruz Biotechnology; Texas, USA).

Techniques: Expressing, Cell Culture, Activity Assay, Control, Isolation, Staining, Transfection, shRNA, Flow Cytometry

Journal: International Journal of Biological Sciences

Article Title: Inhibition of α4β1 Integrin Activity by Small Tellurium Compounds Regulates PD-L1 Expression and Enhances Antitumor Effects

doi: 10.7150/ijbs.95350

Figure Lengend Snippet: SAS inhibits both PD-L1 expression and VLA-4 activity in U937 myelomonocytic human leukemic cells. ( A ) U937 human cells were cultured in the presence or absence of SAS. The cells were collected and stained with either PE-conjugated anti-human PD-L1 antibodies or their respective isotype-matched controls. The results show one representative experiment of 3 performed. ( B ) U937 cells were cultured on VCAM-1- or BSA-coated plates with or without SAS for 1 h. The cells were subsequently washed twice. The percentage of attached cells (representing VLA-4 activity) was determined by the XTT viability test relative to the control PBS. * p<0.001 vs. BSA **p<0.001 vs. PBS. Significance was calculated via one-way ANOVA. The results represent the mean +SE from 3 experiments ( B ). ( C ) The VLA-4 conformational structure was detected by FRET. VLA-4 activation involves the separation of the fluorescently tagged cytoplasmic ends of the α- and β-subunits of the integrin, resulting in a reduction in the FRET signal. U937 cells were transfected with α4-mCFP and β1-mYFP. After 48 hours, the cells were activated and fixed. FRET of the molecular interaction between the cytoplasmic domains of α4 and β1 was determined as described in the Materials and Methods. ( D ) Results for FRET positive controls (cells expressing CFP and YFP encoded on the same plasmid (i.e., maximal FRET obtained in this system) and negative controls (CFP and YFP expressed by different plasmids and undergoing minimal FRET, i.e., only that produced by random colocalizations). *p<0.01 vs. the negative control. Significance was calculated via a two-tailed t test. PLL (poly-L-lysine). FN (Fibronectin).

Techniques: Expressing, Activity Assay, Cell Culture, Staining, Control, Activation Assay, Transfection, Plasmid Preparation, Produced, Negative Control, Two Tailed Test

Journal: International Journal of Biological Sciences

Article Title: Inhibition of α4β1 Integrin Activity by Small Tellurium Compounds Regulates PD-L1 Expression and Enhances Antitumor Effects

doi: 10.7150/ijbs.95350

Figure Lengend Snippet: Associations between VLA-4, IL-10 and PD-L1. ( A ) B16/F10 cells were cultured on FN-coated plates in the presence or absence of SAS at various concentrations. The cells were collected, fixed, permeabilized and stained with PE-conjugated αIL-10 Abs. ( B ) SK-MEL23 cells were cultured on FN-coated plates in the presence or absence of SAS at various concentrations, with or without IFNγ (2 μg/ml), for 48 h. The supernatants were collected, and IL-10 levels were measured by ELISA. *p<0.0001 vs. without IFNγ; **p<0.001 vs. PBS. Significance was calculated via two-way ANOVA. The results represent the mean+SE of 3 experiments. SK-MEL23 cells without ( C, E ) or with IFNγ ( D, F ) were cultured on FN-coated plates in the presence or absence of SAS at various concentrations for 24 h, after which the cells were collected and stained for PD-L1 ( C, D ) or VLA-4 ( E, F ). All FACS results show one representative of 3 experiments. ( G ) SK-MEL23 cells were cultured in the presence or absence of SAS at various concentrations with or without IFNγ (2 μg/ml) for 24 h. The cells were detached and replated again on VCAM-1- or BSA-coated plates with or without SAS at various concentrations for 1 h. The cells were subsequently washed twice . The percentage of attached cells (representing VLA-4 activity) was determined by the XTT viability test relative to the control. *p<0.0001 vs. without IFNγ; **p<0.001 vs. PBS. Significance was calculated via two-way ANOVA. The results represent the mean+SE from 3 experiments. VLA-4 (Very late antigen-4/integrin α4β1). IFNγ (interferon gamma).

Techniques: Cell Culture, Staining, Enzyme-linked Immunosorbent Assay, Activity Assay, Control

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