human il8 (Miltenyi Biotec)

Miltenyi Biotec is a verified supplier

Miltenyi Biotec manufactures this product

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Miltenyi Biotec human il8

A Representative images of wound closure assays of MCF7 cells (WT) after 24 h of treatment with PBS (vehicle control; −) or <t>IL8</t> (250 ng/ml). B Quantification of the extent of wound closure calculated by analyzing the scratched area covered by cells in ( A ) after 24 h using ImageJ software. C Representative images of wound closure assays of Shp1-KO cells (Shp1-KO) after 24 h of treatment with PBS (vehicle control; −) or IL8 (250 ng/ml). D Quantification of the extent of wound closure calculated by analyzing the scratched area covered by cells in ( C ) after 24 h using ImageJ software. E Quantification of Matrigel invasion assays of MCF7 cells treated as in ( A ). F Quantification of Matrigel invasion assays of MCF7 Shp1-KO cells treated as in ( C ). Invasion was quantified by crystal violet staining, elution of invading cells, and data are presented as fold increase in invading cells. Data are mean ± SD of three independent experiments performed in triplicate. **** P < 0.0001, * P < 0.05 versus vehicle control (Student’s t-tests). G Representative images of MCF7 and MCF7 Shp1-KO cells treated with PBS (vehicle control; −) or IL8 (250 ng/ml) for 1 h (as indicated). Cells were fixed and stained with phalloidin (gray) and Hoechst 33342 (blue). Scale bars, 10 μm. H Quantification of ruffle area/cell (expressed as percentage of control) of cells in ( G ) (see “Materials and methods”). Data are mean ± SD of three independent experiments. **** P < 0.0001 versus Ctrl (WT cells treated with vehicle control) (Student’s t-tests).

Human Il8, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+il8/Human+IL-8%2C+premium+grade/pmc13039404-361-2-5
Average 94 stars, based on 2 article reviews

human il8 - by Bioz Stars, 2026-09

94/100 stars

Images

1) Product Images from "Shp1 phosphatase regulates CXCR2 protein stability and IL8-mediated invasiveness in breast cancer"

Article Title: Shp1 phosphatase regulates CXCR2 protein stability and IL8-mediated invasiveness in breast cancer

Journal: Cell Death & Disease

doi: 10.1038/s41419-026-08516-4

A Representative images of wound closure assays of MCF7 cells (WT) after 24 h of treatment with PBS (vehicle control; −) or IL8 (250 ng/ml). B Quantification of the extent of wound closure calculated by analyzing the scratched area covered by cells in ( A ) after 24 h using ImageJ software. C Representative images of wound closure assays of Shp1-KO cells (Shp1-KO) after 24 h of treatment with PBS (vehicle control; −) or IL8 (250 ng/ml). D Quantification of the extent of wound closure calculated by analyzing the scratched area covered by cells in ( C ) after 24 h using ImageJ software. E Quantification of Matrigel invasion assays of MCF7 cells treated as in ( A ). F Quantification of Matrigel invasion assays of MCF7 Shp1-KO cells treated as in ( C ). Invasion was quantified by crystal violet staining, elution of invading cells, and data are presented as fold increase in invading cells. Data are mean ± SD of three independent experiments performed in triplicate. **** P < 0.0001, * P < 0.05 versus vehicle control (Student’s t-tests). G Representative images of MCF7 and MCF7 Shp1-KO cells treated with PBS (vehicle control; −) or IL8 (250 ng/ml) for 1 h (as indicated). Cells were fixed and stained with phalloidin (gray) and Hoechst 33342 (blue). Scale bars, 10 μm. H Quantification of ruffle area/cell (expressed as percentage of control) of cells in ( G ) (see “Materials and methods”). Data are mean ± SD of three independent experiments. **** P < 0.0001 versus Ctrl (WT cells treated with vehicle control) (Student’s t-tests).

Figure Legend Snippet: A Representative images of wound closure assays of MCF7 cells (WT) after 24 h of treatment with PBS (vehicle control; −) or IL8 (250 ng/ml). B Quantification of the extent of wound closure calculated by analyzing the scratched area covered by cells in ( A ) after 24 h using ImageJ software. C Representative images of wound closure assays of Shp1-KO cells (Shp1-KO) after 24 h of treatment with PBS (vehicle control; −) or IL8 (250 ng/ml). D Quantification of the extent of wound closure calculated by analyzing the scratched area covered by cells in ( C ) after 24 h using ImageJ software. E Quantification of Matrigel invasion assays of MCF7 cells treated as in ( A ). F Quantification of Matrigel invasion assays of MCF7 Shp1-KO cells treated as in ( C ). Invasion was quantified by crystal violet staining, elution of invading cells, and data are presented as fold increase in invading cells. Data are mean ± SD of three independent experiments performed in triplicate. **** P < 0.0001, * P < 0.05 versus vehicle control (Student’s t-tests). G Representative images of MCF7 and MCF7 Shp1-KO cells treated with PBS (vehicle control; −) or IL8 (250 ng/ml) for 1 h (as indicated). Cells were fixed and stained with phalloidin (gray) and Hoechst 33342 (blue). Scale bars, 10 μm. H Quantification of ruffle area/cell (expressed as percentage of control) of cells in ( G ) (see “Materials and methods”). Data are mean ± SD of three independent experiments. **** P < 0.0001 versus Ctrl (WT cells treated with vehicle control) (Student’s t-tests).

Techniques Used: Control, Software, Staining

$A Representative western blot using anti-phosphoserine 591 in Shp1 (pSer591-Shp1) specific antibody in MCF7 cells treated with PBS (vehicle control; −) or IL8 (250 ng/ml; +) for the indicated times. Total Shp1 was used as loading controls. Data are representative of five independent experiments. Molecular weight standards (kDa) are indicated on the left of each panel. B Densitometric quantification of the blot in ( A ) (60 min). Data are mean ± SD of five independent experiments. * P < 0.05 versus control. C Schematic representation of the experimental workflow for Shp1 immunoprecipitation and phosphatase activity assay. D Representative immunoprecipitated Shp1 fraction (IP) from MCF7 cells treated with PBS (vehicle control; −) or IL8 (250 ng/ml) for 60 min (as indicated). The top panels show the input and the unbound fractions. Molecular weight standards (kDa) are indicated on the left of each panel. E Quantification of enzymatic activity of Shp1 immunoprecipitated as in ( C ) (see “Materials and methods”). Data are mean ± SD of five independent experiments. **** P < 0.0001 versus ctrl (WT cells treated with vehicle control) (Student’s t-tests). F Representative western blot using anti-phosphoserine 591 in Shp1 (pSer591-Shp1) specific antibody in MCF7 cells treated with PBS (vehicle control; −) or IL8 (250 ng/ml; +) for 1 h in absence or presence of PKCi (1 μM). Total Shp1 and GAPDH were used as loading controls. Data are representative of at least three independent experiments. Molecular weight standards (kDa) are indicated on the left of each panel. G Densitometric quantification of the blot in ( F ). Data are mean ± SD of at least three independent experiments. * P < 0.05 versus ctrl (vehicle control; −).$
Figure Legend Snippet: A Representative western blot using anti-phosphoserine 591 in Shp1 (pSer591-Shp1) specific antibody in MCF7 cells treated with PBS (vehicle control; −) or IL8 (250 ng/ml; +) for the indicated times. Total Shp1 was used as loading controls. Data are representative of five independent experiments. Molecular weight standards (kDa) are indicated on the left of each panel. B Densitometric quantification of the blot in ( A ) (60 min). Data are mean ± SD of five independent experiments. * P < 0.05 versus control. C Schematic representation of the experimental workflow for Shp1 immunoprecipitation and phosphatase activity assay. D Representative immunoprecipitated Shp1 fraction (IP) from MCF7 cells treated with PBS (vehicle control; −) or IL8 (250 ng/ml) for 60 min (as indicated). The top panels show the input and the unbound fractions. Molecular weight standards (kDa) are indicated on the left of each panel. E Quantification of enzymatic activity of Shp1 immunoprecipitated as in ( C ) (see “Materials and methods”). Data are mean ± SD of five independent experiments. **** P < 0.0001 versus ctrl (WT cells treated with vehicle control) (Student’s t-tests). F Representative western blot using anti-phosphoserine 591 in Shp1 (pSer591-Shp1) specific antibody in MCF7 cells treated with PBS (vehicle control; −) or IL8 (250 ng/ml; +) for 1 h in absence or presence of PKCi (1 μM). Total Shp1 and GAPDH were used as loading controls. Data are representative of at least three independent experiments. Molecular weight standards (kDa) are indicated on the left of each panel. G Densitometric quantification of the blot in ( F ). Data are mean ± SD of at least three independent experiments. * P < 0.05 versus ctrl (vehicle control; −).

Techniques Used: Western Blot, Control, Molecular Weight, Immunoprecipitation, Phosphatase Assay, Activity Assay

Representative western blot using anti-phosphotyrosine 307 in PP2A (pTyr307-PP2A), anti-phosphoserine 347 in CXCR2 (pSer347-CXCR2), and anti-phosphoserine 591 in Shp1 (pSer591-Shp1) specific antibodies in MCF7 ( A ) and MCF7 Shp1-KO ( C ) cells treated with PBS (vehicle control; −) or IL8 (250 ng/ml; +) for 1 h. Total CXCR2, PP2A, Shp1, β-tubulin, and vinculin were used as loading controls. Data are representative of at least three independent experiments. Molecular weight standards (kDa) are indicated on the left of each panel. B Densitometric quantification of the blot in ( A ). Data are mean ± SD of at least three independent experiments. * P < 0.05 versus ctrl. D Representative western blot using anti-CXCR2, anti-phosphoserine 347 in CXCR2 (pSer347-CXCR2) and anti-CXCR1 specific antibodies in cell lysates from MCF7 WT and Shp1-KO cells. GAPDH was used as loading control. Molecular weight standards (kDa) are indicated on the left of each panel. Data are representative of three independent experiments. E , F Densitometric quantification of the blots in ( D ). *** P < 0.0005; ** P < 0.005 versus WT cells (Student’s t-tests). G Analysis by qRT-PCR of mRNA levels of CXCR2 in MCF7 WT and Shp1-KO cells. β-actin was used as housekeeping gene. Data are expressed as log2 fold change of Shp1-KO versus WT cells. Data are means ± SD of three independent experiments performed in triplicate. H Representative western blot using anti-CXCR2 specific antibody in MCF7 cells treated with PBS (vehicle control; −) or SSG (10 μM) for 48 h. Vinculin was used as loading control. Data are representative of three independent experiments. Molecular weight standards (kDa) are indicated on the left of each panel. I Densitometric quantification of the blot in ( H ). * P < 0.05 versus Ctrl (Student’s t-tests). J Representative western blot using anti-CXCR2 and anti-Shp1 specific antibodies in MCF7 Shp1-KO cells transfected with Shp1 WT and Shp1 C455S mutant. The expression levels of Shp1 WT and Shp1 C455S indicate comparable amounts of proteins. Vinculin was used as loading control. Data are representative of five independent experiments. Molecular weight standards (kDa) are indicated on the left of each panel. K Densitometric quantification of the blot in ( J ). * P < 0.05 versus Ctrl (untransfected cells) (Student’s t-tests).

Figure Legend Snippet: Representative western blot using anti-phosphotyrosine 307 in PP2A (pTyr307-PP2A), anti-phosphoserine 347 in CXCR2 (pSer347-CXCR2), and anti-phosphoserine 591 in Shp1 (pSer591-Shp1) specific antibodies in MCF7 ( A ) and MCF7 Shp1-KO ( C ) cells treated with PBS (vehicle control; −) or IL8 (250 ng/ml; +) for 1 h. Total CXCR2, PP2A, Shp1, β-tubulin, and vinculin were used as loading controls. Data are representative of at least three independent experiments. Molecular weight standards (kDa) are indicated on the left of each panel. B Densitometric quantification of the blot in ( A ). Data are mean ± SD of at least three independent experiments. * P < 0.05 versus ctrl. D Representative western blot using anti-CXCR2, anti-phosphoserine 347 in CXCR2 (pSer347-CXCR2) and anti-CXCR1 specific antibodies in cell lysates from MCF7 WT and Shp1-KO cells. GAPDH was used as loading control. Molecular weight standards (kDa) are indicated on the left of each panel. Data are representative of three independent experiments. E , F Densitometric quantification of the blots in ( D ). *** P < 0.0005; ** P < 0.005 versus WT cells (Student’s t-tests). G Analysis by qRT-PCR of mRNA levels of CXCR2 in MCF7 WT and Shp1-KO cells. β-actin was used as housekeeping gene. Data are expressed as log2 fold change of Shp1-KO versus WT cells. Data are means ± SD of three independent experiments performed in triplicate. H Representative western blot using anti-CXCR2 specific antibody in MCF7 cells treated with PBS (vehicle control; −) or SSG (10 μM) for 48 h. Vinculin was used as loading control. Data are representative of three independent experiments. Molecular weight standards (kDa) are indicated on the left of each panel. I Densitometric quantification of the blot in ( H ). * P < 0.05 versus Ctrl (Student’s t-tests). J Representative western blot using anti-CXCR2 and anti-Shp1 specific antibodies in MCF7 Shp1-KO cells transfected with Shp1 WT and Shp1 C455S mutant. The expression levels of Shp1 WT and Shp1 C455S indicate comparable amounts of proteins. Vinculin was used as loading control. Data are representative of five independent experiments. Molecular weight standards (kDa) are indicated on the left of each panel. K Densitometric quantification of the blot in ( J ). * P < 0.05 versus Ctrl (untransfected cells) (Student’s t-tests).

Techniques Used: Western Blot, Control, Molecular Weight, Quantitative RT-PCR, Transfection, Mutagenesis, Expressing

A Representative confocal microscopy images of MCF7 (WT) and MCF7 Shp1-KO cells (Shp1-KO). Cells were fixed and stained with Hoechst 33342 (nuclei, blue), WGA (plasma membrane, red), and an anti-CXCR2 antibody (endogenous CXCR2; green). Insets, right: Magnification of a plasma membrane section. Scale bars, 10 μm. B Pearson correlation coefficient of CXCR2 co-localization with WGA in ( A ). Data are means ± SD of three independent experiments. **** P < 0.0001 versus WT (Student’s t-tests). C Working model of CXCR2 trafficking and degradation under normal versus prolonged IL8 stimulation. Under normal stimulation ( left panel ), IL8 binding to CXCR2 activates downstream G-protein signaling (Gα/βγ). The receptor is subsequently phosphorylated by GPCR kinases (GRKs), leading to β-arrestin recruitment and receptor internalization. Internalized CXCR2 is either directed to recycling pathways back to the plasma membrane or targeted for degradation. Receptor fate is determined by the balance between phosphorylation and dephosphorylation, the latter mediated by active PP2A. Active PP2A promotes receptor recycling by removing phosphate groups from internalized CXCR2. Under prolonged stimulation ( right panel ), PKC-mediated inhibition of Shp1 phosphatase leads to inhibition of PP2A activity, preventing receptor dephosphorylation. This shift favors CXCR2 ubiquitination, leading to increased receptor degradation over recycling. This contributes to receptor desensitization and downregulation at the cell surface.

Figure Legend Snippet: A Representative confocal microscopy images of MCF7 (WT) and MCF7 Shp1-KO cells (Shp1-KO). Cells were fixed and stained with Hoechst 33342 (nuclei, blue), WGA (plasma membrane, red), and an anti-CXCR2 antibody (endogenous CXCR2; green). Insets, right: Magnification of a plasma membrane section. Scale bars, 10 μm. B Pearson correlation coefficient of CXCR2 co-localization with WGA in ( A ). Data are means ± SD of three independent experiments. **** P < 0.0001 versus WT (Student’s t-tests). C Working model of CXCR2 trafficking and degradation under normal versus prolonged IL8 stimulation. Under normal stimulation ( left panel ), IL8 binding to CXCR2 activates downstream G-protein signaling (Gα/βγ). The receptor is subsequently phosphorylated by GPCR kinases (GRKs), leading to β-arrestin recruitment and receptor internalization. Internalized CXCR2 is either directed to recycling pathways back to the plasma membrane or targeted for degradation. Receptor fate is determined by the balance between phosphorylation and dephosphorylation, the latter mediated by active PP2A. Active PP2A promotes receptor recycling by removing phosphate groups from internalized CXCR2. Under prolonged stimulation ( right panel ), PKC-mediated inhibition of Shp1 phosphatase leads to inhibition of PP2A activity, preventing receptor dephosphorylation. This shift favors CXCR2 ubiquitination, leading to increased receptor degradation over recycling. This contributes to receptor desensitization and downregulation at the cell surface.

Techniques Used: Confocal Microscopy, Staining, Clinical Proteomics, Membrane, Binding Assay, Phospho-proteomics, De-Phosphorylation Assay, Inhibition, Activity Assay, Ubiquitin Proteomics

$A Representative western blot using anti-CXCR2 specific antibody in MCF7 (WT) and MCF7 Shp1-KO (Shp1-KO) cells treated with CHX (25 μg/ml) and IL8 (250 ng/ml) for the indicated time points (see “Materials and methods”). β-tubulin was used as loading control. Data are representative of at least five independent experiments. Molecular weight standards (kDa) are indicated on the left of each panel. B Densitometric quantification of the blot in ( A ). ** P < 0.005 versus time zero (Student’s t-tests). Representative western blot using anti-CXCR2 specific antibody in MCF7 Shp1-KO cells treated with DMSO (vehicle control; Ctrl), 20 μM ALLN, 25 μM MG132, or 1 μM Bortezomib for 4 h ( C ) or with DMSO (vehicle control; Ctrl), 0.1 and 0.3 μM Bafilomycin A1 or 50 and 100 μM Chloroquine for 4 h ( E ). The expression levels of p21 and LC3 I-II were monitored to evaluate inhibitors efficacy. β-tubulin was used as loading control. Data are representative of at least three independent experiments. Molecular weight standards (kDa) are indicated on the left of each panel. D Densitometric quantification of the blot in ( C ). * P < 0.05 versus Ctrl (Student’s t-tests). F Representative western blot using anti-ubiquitin antibody in the precipitate CXCR2 fraction (IP) from MCF7 cells transfected with an empty 3xFlag-vector or with 3xFlag-CXCR2 and from MCF7 Shp1-KO cells transfected with 3xFlag-CXCR2. The cells were treated with Bortezomib (1 μM) for 16 h, and then the lysates were immunoprecipitated using anti-Flag beads (see “Materials and methods”). Ubiquitin staining of 3xFlag-CXCR2 is shown with two different exposure times (low and high) for better interpretation. The protein levels of total 3xFlag-CXCR2 were detected with anti-Flag antibody and indicate comparable amounts of transfected protein (lower panel). Data are representative of six independent experiments. Molecular weight standards (kDa) are indicated on the left of each panel. G Quantification of ubiquitinylated receptor as in ( F ). * P < 0.05 versus WT (MCF7 cells transfected with 3xFlag-CXCR2) (Student’s t-tests). H Representative western blot using anti-ubiquitin antibody in the precipitate CXCR2 fraction (IP) from MCF7 cells transfected with an empty 3xFlag-vector or with 3xFlag-CXCR2 (WT) or with 3xFlag-CXCR2 S347A mutant (S347A). The cells were treated with Bortezomib (1 μM) for 16 h, and then the lysates were immunoprecipitated using anti-Flag beads (see “Materials and methods”). The protein levels of total 3xFlag-CXCR2 were detected with anti-Flag antibody and indicate comparable amounts of transfected proteins (lower panel). Data are representative of four independent experiments. Molecular weight standards (kDa) are indicated on the left of each panel. I Quantification of ubiquitinylated receptor as in ( H ). * P < 0.05 versus CXCR2 (MCF7 cells transfected with 3xFlag-CXCR2) (Student’s t-tests). J Representative flow cytometry analysis of CXCR2-surface expression of MCF7 (WT) and MCF7 Shp1-KO (Shp1-KO) cells (as indicated) stained with PerCP-Cy5.5-anti-CXCR2 antibody. The gate was set according to the negative control in the different cell lines. Data are representative of at least four independent experiments. K Quantification of surface expression of CXCR2 in MCF7 WT and Shp1-KO cells treated with IL8 (250 ng/ml) for the indicated time points and analyzed by flow cytometry (see “Materials and methods”). Data are expressed as percentages of control (untreated MCF7 WT cells). Data are mean ± SE of at least four independent experiments.$
Figure Legend Snippet: A Representative western blot using anti-CXCR2 specific antibody in MCF7 (WT) and MCF7 Shp1-KO (Shp1-KO) cells treated with CHX (25 μg/ml) and IL8 (250 ng/ml) for the indicated time points (see “Materials and methods”). β-tubulin was used as loading control. Data are representative of at least five independent experiments. Molecular weight standards (kDa) are indicated on the left of each panel. B Densitometric quantification of the blot in ( A ). ** P < 0.005 versus time zero (Student’s t-tests). Representative western blot using anti-CXCR2 specific antibody in MCF7 Shp1-KO cells treated with DMSO (vehicle control; Ctrl), 20 μM ALLN, 25 μM MG132, or 1 μM Bortezomib for 4 h ( C ) or with DMSO (vehicle control; Ctrl), 0.1 and 0.3 μM Bafilomycin A1 or 50 and 100 μM Chloroquine for 4 h ( E ). The expression levels of p21 and LC3 I-II were monitored to evaluate inhibitors efficacy. β-tubulin was used as loading control. Data are representative of at least three independent experiments. Molecular weight standards (kDa) are indicated on the left of each panel. D Densitometric quantification of the blot in ( C ). * P < 0.05 versus Ctrl (Student’s t-tests). F Representative western blot using anti-ubiquitin antibody in the precipitate CXCR2 fraction (IP) from MCF7 cells transfected with an empty 3xFlag-vector or with 3xFlag-CXCR2 and from MCF7 Shp1-KO cells transfected with 3xFlag-CXCR2. The cells were treated with Bortezomib (1 μM) for 16 h, and then the lysates were immunoprecipitated using anti-Flag beads (see “Materials and methods”). Ubiquitin staining of 3xFlag-CXCR2 is shown with two different exposure times (low and high) for better interpretation. The protein levels of total 3xFlag-CXCR2 were detected with anti-Flag antibody and indicate comparable amounts of transfected protein (lower panel). Data are representative of six independent experiments. Molecular weight standards (kDa) are indicated on the left of each panel. G Quantification of ubiquitinylated receptor as in ( F ). * P < 0.05 versus WT (MCF7 cells transfected with 3xFlag-CXCR2) (Student’s t-tests). H Representative western blot using anti-ubiquitin antibody in the precipitate CXCR2 fraction (IP) from MCF7 cells transfected with an empty 3xFlag-vector or with 3xFlag-CXCR2 (WT) or with 3xFlag-CXCR2 S347A mutant (S347A). The cells were treated with Bortezomib (1 μM) for 16 h, and then the lysates were immunoprecipitated using anti-Flag beads (see “Materials and methods”). The protein levels of total 3xFlag-CXCR2 were detected with anti-Flag antibody and indicate comparable amounts of transfected proteins (lower panel). Data are representative of four independent experiments. Molecular weight standards (kDa) are indicated on the left of each panel. I Quantification of ubiquitinylated receptor as in ( H ). * P < 0.05 versus CXCR2 (MCF7 cells transfected with 3xFlag-CXCR2) (Student’s t-tests). J Representative flow cytometry analysis of CXCR2-surface expression of MCF7 (WT) and MCF7 Shp1-KO (Shp1-KO) cells (as indicated) stained with PerCP-Cy5.5-anti-CXCR2 antibody. The gate was set according to the negative control in the different cell lines. Data are representative of at least four independent experiments. K Quantification of surface expression of CXCR2 in MCF7 WT and Shp1-KO cells treated with IL8 (250 ng/ml) for the indicated time points and analyzed by flow cytometry (see “Materials and methods”). Data are expressed as percentages of control (untreated MCF7 WT cells). Data are mean ± SE of at least four independent experiments.

Techniques Used: Western Blot, Control, Molecular Weight, Expressing, Ubiquitin Proteomics, Transfection, Plasmid Preparation, Immunoprecipitation, Staining, Mutagenesis, Flow Cytometry, Negative Control

Quantification of Matrigel invasion assays of T47D ( A ), MDA-MB-231 ( B ), BT-549 ( C ), SK-BR-3 ( D ), and MDA-MB-453 ( E ) cells after 24 h of treatment with PBS (vehicle control; −) or IL8 (250 ng/ml) in absence or presence of SSG (10 μM) (as indicated). Invasion was quantified by crystal violet staining, elution of invading cells, and data are presented as fold increase in invading cells. Data are mean ± SE of two independent experiments. **** P < 0.0001, *** P < 0.0005, ** P < 0.005, * P < 0.05 versus Ctrl (vehicle control, PBS) (Student’s t-tests). Representative western blot using anti-phosphoserine 591 in Shp1 (pSer591-Shp1), anti-phosphotyrosine 307 in PP2A (pTyr307-PP2A) and anti-phosphoserine 347 in CXCR2 (pSer347-CXCR2) in T47D ( F ), MDA-MB-231 ( G ), BT-549 ( H ), SK-BR-3 ( I ) and MDA-MB-453 ( J ) cells treated with PBS (vehicle control; −) or IL8 (250 ng/ml; +) for 1 h. Total Shp1, PP2A, and CXCR2 were used as loading controls. Data are representative of at least three independent experiments. Molecular weight standards (kDa) are indicated on the left of each panel. K – M Densitometric quantification of the blots in ( F – H ). **** P < 0.0001, *** P < 0.0005, ** P < 0.005, * P < 0.05 versus Ctrl (vehicle control, PBS) (Student’s t-tests).

Figure Legend Snippet: Quantification of Matrigel invasion assays of T47D ( A ), MDA-MB-231 ( B ), BT-549 ( C ), SK-BR-3 ( D ), and MDA-MB-453 ( E ) cells after 24 h of treatment with PBS (vehicle control; −) or IL8 (250 ng/ml) in absence or presence of SSG (10 μM) (as indicated). Invasion was quantified by crystal violet staining, elution of invading cells, and data are presented as fold increase in invading cells. Data are mean ± SE of two independent experiments. **** P < 0.0001, *** P < 0.0005, ** P < 0.005, * P < 0.05 versus Ctrl (vehicle control, PBS) (Student’s t-tests). Representative western blot using anti-phosphoserine 591 in Shp1 (pSer591-Shp1), anti-phosphotyrosine 307 in PP2A (pTyr307-PP2A) and anti-phosphoserine 347 in CXCR2 (pSer347-CXCR2) in T47D ( F ), MDA-MB-231 ( G ), BT-549 ( H ), SK-BR-3 ( I ) and MDA-MB-453 ( J ) cells treated with PBS (vehicle control; −) or IL8 (250 ng/ml; +) for 1 h. Total Shp1, PP2A, and CXCR2 were used as loading controls. Data are representative of at least three independent experiments. Molecular weight standards (kDa) are indicated on the left of each panel. K – M Densitometric quantification of the blots in ( F – H ). **** P < 0.0001, *** P < 0.0005, ** P < 0.005, * P < 0.05 versus Ctrl (vehicle control, PBS) (Student’s t-tests).

Techniques Used: Control, Staining, Western Blot, Molecular Weight

A The plot shows the number of the differentially expressed genes (DEG) identified between the contrasts “luminal vs TNBC,” “luminal vs HER2” cell lines, and “TNBC vs HER2” cell lines defined using adjusted p value < 0.05 and fold change ≥2. Note that the numbers on Y axis represent the total number of selected genes found for each contrast. The genes downregulated and upregulated are shown in light blue and black, respectively. Heatmap of significant genes for comparison “luminal vs HER2” cell lines ( B ) and “TNBC vs HER2” cell lines ( C ). The Heatmap shows gene intensity per sample relative to the average level across all samples. Individual genes are shown on the Y axis, while samples are shown along the X axis. Red and blue cells correspond to higher and lower RNA-seq levels, respectively. A maximum of 1000 features and 1000 samples are shown (selected at random when the number exceeds these limits). D , E Dot plot of significantly enriched manually curated Reactome pathways of genes identified in ( B , C ). Enrichment results were plotted as manually curated Reactome pathways (Y axis) against odds ratio (OR, X axis), calculated as the ratio of significant genes observed over that which could be expected by chance. Points are colored by −log10(adjusted P value) value and sized by the number of member genes within each manually curated Reactome pathway. Boxplots of expression of Shp1 ( F ), CXCR2 ( G ), and IL8 ( H ) across all samples grouped by breast cancer cell lines. Each dot represents a sample and expression (Y axis) has been summarized by tissue type (X axis). Expression is presented as Z-scores of the TMM normalized values. *** P < 0.005, ** P < 0.005; * P < 0.05 (One-way analysis). Analysis by qRT-PCR of mRNA levels of Shp1 ( I ), CXCR2 ( J ), and IL8 ( K ) in MCF7, SK-BR-3, and MDA-MB-231 cells. GAPDH was used as housekeeping gene. Data are means ± SE of three independent experiments performed in triplicate. **** P < 0.0001, *** P < 0.0005, ** P < 0.005 (One-way analysis).

Figure Legend Snippet: A The plot shows the number of the differentially expressed genes (DEG) identified between the contrasts “luminal vs TNBC,” “luminal vs HER2” cell lines, and “TNBC vs HER2” cell lines defined using adjusted p value < 0.05 and fold change ≥2. Note that the numbers on Y axis represent the total number of selected genes found for each contrast. The genes downregulated and upregulated are shown in light blue and black, respectively. Heatmap of significant genes for comparison “luminal vs HER2” cell lines ( B ) and “TNBC vs HER2” cell lines ( C ). The Heatmap shows gene intensity per sample relative to the average level across all samples. Individual genes are shown on the Y axis, while samples are shown along the X axis. Red and blue cells correspond to higher and lower RNA-seq levels, respectively. A maximum of 1000 features and 1000 samples are shown (selected at random when the number exceeds these limits). D , E Dot plot of significantly enriched manually curated Reactome pathways of genes identified in ( B , C ). Enrichment results were plotted as manually curated Reactome pathways (Y axis) against odds ratio (OR, X axis), calculated as the ratio of significant genes observed over that which could be expected by chance. Points are colored by −log10(adjusted P value) value and sized by the number of member genes within each manually curated Reactome pathway. Boxplots of expression of Shp1 ( F ), CXCR2 ( G ), and IL8 ( H ) across all samples grouped by breast cancer cell lines. Each dot represents a sample and expression (Y axis) has been summarized by tissue type (X axis). Expression is presented as Z-scores of the TMM normalized values. *** P < 0.005, ** P < 0.005; * P < 0.05 (One-way analysis). Analysis by qRT-PCR of mRNA levels of Shp1 ( I ), CXCR2 ( J ), and IL8 ( K ) in MCF7, SK-BR-3, and MDA-MB-231 cells. GAPDH was used as housekeeping gene. Data are means ± SE of three independent experiments performed in triplicate. **** P < 0.0001, *** P < 0.0005, ** P < 0.005 (One-way analysis).

Techniques Used: Comparison, RNA Sequencing, Expressing, Quantitative RT-PCR

Control:

Article Title: Shp1 phosphatase regulates CXCR2 protein stability and IL8-mediated invasiveness in breast cancer
Article Snippet: The antibodies used were: polyclonal anti-CXCR2 (PA5-102662), polyclonal anti-pSer347-CXCR2 (PA5-104850), PerCP-conjugated anti-CXCR2 (46-1829-42), monoclonal anti-PP2Aα (MA5-18060), polyclonal anti-Tyr307-PP2Aα (PA5-36874) and Alexa Fluor 555-conjugated WGA ( W32464 ) from Invitrogen (Thermo Fisher Scientific, Waltham, MA, USA), monoclonal anti-Shp1 (26516), polyclonal anti-Vinculin (4650), monoclonal anti-GAPDH (97166) monoclonal anti-LC3 (12741), monoclonal anti-Ubiquitin (20326), monoclonal anti-p21 (2947), anti-rabbit IgG/HRP-linked secondary antibody (7074) and anti-mouse IgG/HRP-linked secondary antibody (7076) from Cell Signaling Technology (Danvers, MA, USA), polyclonal anti-pSer591-Shp1 (Ab41436, Abcam, Cambridge, UK), monoclonal anti-β-Tubulin (E-AB-20033, Elabscience, Houston TX, USA).anti-p21 (2947), anti-rabbit IgG/HRP-linked secondary antibody (7074) and anti-mouse IgG/HRP-linked secondary antibody (7076) from Cell Signaling Technology (Danvers, MA, USA), polyclonal anti-pSer591-Shp1 (Ab41436, Abcam, Cambridge, UK), monoclonal anti-β-Tubulin (E-AB-20033, Elabscience, Houston TX, USA). ... Reagents included human IL8 (130-122-360, Miltenyi Biotec, Bergisch Gladbach, Germany), SSG inhibitor Sodium Stibogluconate (567565), Chloroquine diphosphate salt (C6628), PKCi Bisindolylmaleimide I (203290) and Cycloheximide (01810) from Merck Millipore (Burlington, MA, USA), MG132 (2194) and Bafilomycin A1 (54645) from Cell Signaling Technology, ALLN (208719, Calbiochem, EMD Chemicals, San Diego, CA, USA), Bortezomib (sc-217785, Santa Cruz Biotechnology, Dallas, TX, USA).. Expression vectors used were Shp1 and Shp1-C455S (kindly provided by F.D.Expression vectors used were Shp1 and Shp1-C455S (kindly provided by F.D.

Article Title: Shp1 phosphatase regulates CXCR2 protein stability and IL8-mediated invasiveness in breast cancer.
Article Snippet: Antibodies, reagents and constructs The antibodies used were: polyclonal anti-CXCR2 (PA5-102662), polyclonal anti-pSer347- CXCR2 (PA5-104850), PerCP-conjugated anti-CXCR2 (46-1829-42), monoclonal anti-PP2A (MA5-18060), polyclonal anti-Tyr307-PP2A (PA5-36874) and Alexa Fluor 555-conjugated WGA (W32464) from Invitrogen (Thermo Fisher Scientific, Waltham, MA, USA), monoclonal anti-Shp1 (26516), polyclonal anti-Vinculin (4650), monoclonal anti-GAPDH (97166) monoclonal anti-LC3 (12741), monoclonal anti-Ubiquitin (20326), monoclonal anti-p21 (2947), anti-rabbit IgG/HRP-linked secondary antibody (7074) and anti-mouse IgG/HRP-linked secondary antibody (7076) from Cell Signaling Technology (Danvers, MA, USA), polyclonal anti- pSer591-Shp1 (Ab41436, Abcam, Cambridge, UK), monoclonal anti--Tubulin (E-AB-20033, Elabscience, Houston TX, USA).(2947), anti-rabbit IgG/HRP-linked secondary antibody (7074) and anti-mouse IgG/HRP-linked secondary antibody (7076) from Cell Signaling Technology (Danvers, MA, USA), polyclonal anti- pSer591-Shp1 (Ab41436, Abcam, Cambridge, UK), monoclonal anti--Tubulin (E-AB-20033, Elabscience, Houston TX, USA). ... Reagents included human IL8 (130-122-360, Miltenyi Biotec, Bergisch Gladbach, Germany), SSG inhibitor Sodium Stibogluconate (567565), Chloroquine diphosphate salt (C6628), PKCi Bisindolylmaleimide I (203290) and Cycloheximide (01810) from Merck Millipore (Burlington, MA, USA), MG132 (2194) and Bafilomycin A1 (54645) from Cell Signaling Technology, ALLN (208719, Calbiochem, EMD Chemicals, San Diego, CA, USA), Bortezomib (sc-217785, Santa Cruz Biotechnology, Dallas, TX, USA).. Expression vectors used were Shp1 and Shp1-C455S (kindly provided by F.D.Expression vectors used were Shp1 and Shp1-C455S (kindly provided by F.D.

Review

human il8 (Miltenyi Biotec)

Structured Review

Images

1) Product Images from "Shp1 phosphatase regulates CXCR2 protein stability and IL8-mediated invasiveness in breast cancer"

Related Articles

Control:

Software:

Staining:

Western Blot:

Molecular Weight:

Immunoprecipitation:

Phosphatase Assay:

Activity Assay:

Quantitative RT-PCR:

Transfection:

Mutagenesis:

Expressing:

Confocal Microscopy:

Clinical Proteomics:

Membrane:

Binding Assay:

Phospho-proteomics:

De-Phosphorylation Assay:

Inhibition:

Ubiquitin Proteomics:

Plasmid Preparation:

Flow Cytometry:

Negative Control:

Comparison:

RNA Sequencing:

Similar Products

Image Search Results