mouse monoclonal anti upa receptor anti upar  (R&D Systems)

Journal: PLoS ONE

Article Title: Plasmin Plays an Essential Role in Amplification of Psoriasiform Skin Inflammation in Mice

doi: 10.1371/journal.pone.0016483

Figure Lengend Snippet: ( A ) A significant reduction in the expression of PLG in PBMC of psoriasis patients was detected by flow cytometry. ( B ) Cryosections from lesional skin of psoriasis patients or normal volunteers were stained with a PLG mAb (red). ( C ) Lysates from lesional skin of psoriasis patients or normal skin of volunteers were subjected to western blotting for PLG expression. Human PLG was used as a positive control (right panel). Actin - loading control. ( D, E ) Cryosections from lesional skin of psoriasis patients or normal volunteers were stained with uPA or tPA mAbs (green) for its expression. Original magnification for immunofluorescent staining, ×400. e, epidermis; d, dermis. Dotted lines indicate the border between epidermis and dermis.

Article Snippet: The following antibodies were used for flow cytometry, all in conjunction with AlexaFluor 555 (Caltag Laboratories): anti-human PLG (Santa Cruz), anti-human uPA, anti-human uPA (R&D), and anti-human annexin A2 (ABGENT).

Techniques: Expressing, Flow Cytometry, Staining, Western Blot, Positive Control

Journal: PLoS ONE

Article Title: Plasmin Plays an Essential Role in Amplification of Psoriasiform Skin Inflammation in Mice

doi: 10.1371/journal.pone.0016483

Figure Lengend Snippet: ( A ) A significant reduction in the expression of PLG in PBMC of psoriasis patients was detected by flow cytometry. ( B ) Cryosections from lesional skin of psoriasis patients or normal volunteers were stained with a PLG mAb (red). ( C ) Lysates from lesional skin of psoriasis patients or normal skin of volunteers were subjected to western blotting for PLG expression. Human PLG was used as a positive control (right panel). Actin - loading control. ( D, E ) Cryosections from lesional skin of psoriasis patients or normal volunteers were stained with uPA or tPA mAbs (green) for its expression. Original magnification for immunofluorescent staining, ×400. e, epidermis; d, dermis. Dotted lines indicate the border between epidermis and dermis.

Article Snippet: The following antibodies were used for flow cytometry, all in conjunction with AlexaFluor 555 (Caltag Laboratories): anti-human PLG (Santa Cruz), anti-human uPA, anti-human uPA (R&D), and anti-human annexin A2 (ABGENT).

Techniques: Expressing, Flow Cytometry, Staining, Western Blot, Positive Control

Journal: BMC Cancer

Article Title: EMMPRIN/CD147 up-regulates urokinase-type plasminogen activator: implications in oral tumor progression

doi: 10.1186/1471-2407-12-115

Figure Lengend Snippet: EMMPRIN and uPA expression in human oral dysplastic and squamous carcinoma lesions . A- EMMPRIN and uPA mRNA expression in oral dysplastic and tumor tissues. RNA was extracted from formalin-fixed paraffin-embedded tissue sections of 6 normal oral mucosa (Normal) , 4 intra epithelial neoplasia ( Dysplastic ), 8 micro-invasive OSCC and 8 invasive OSCC, and were analysed for EMMPRIN and uPA expression using qRT-PCR assays. * denotes significant difference with p < 0.05. B- Immunohistochemical staining of EMMPRIN and uPA in sections of human oral dysplastic, micro-invasive and invasive squamous carcinoma lesions. Tissue sections from patients with oral dysplastic lesions (OIN), micro-invasive lesions (Micro-inv) and invasive oral squamous carcinoma lesions (INV) lesion sections were subjected to single-labeled immunohistochemistry using mouse anti-human EMMPRIN mAb or goat anti-human uPA Ab and counterstained with HES. EMMPRIN and uPA were colocalized, their staining was heterogeneous with particularly high staining at some regions. Staining was more intense in the tumor sections.

Article Snippet: For immunohistochemistry, FFPE sections were stained with a mouse anti-human EMMPRIN antibody (BD-Pharmingen) and a goat anti human uPA antibody (American diagnostica).

Techniques: Expressing, Formalin-fixed Paraffin-Embedded, Quantitative RT-PCR, Immunohistochemical staining, Staining, Labeling, Immunohistochemistry

Journal: BMC Cancer

Article Title: EMMPRIN/CD147 up-regulates urokinase-type plasminogen activator: implications in oral tumor progression

doi: 10.1186/1471-2407-12-115

Figure Lengend Snippet: EMMPRIN regulates uPA in oral dysplastic DOK and tumor SCC-9 cell lines . A, (a) CHO cells were transfected with EMMPRIN cDNA as previously described . Membranes were isolated from CHO-Emp and CHO control cells (CHO-Emp Mb and CHO Mb, respectively) by differential centrifugation and 10 μg of membrane extract were analyzed for EMMPRIN content by immunoblotting. (b) DOK and SCC-9 cells (80% confluence) were incubated with 20 μg/mL CHO Mb or CHO-Emp Mb in serum-free medium for 24 h and the conditioned medium was analyzed for uPA activity using casein-plasminogen zymography and for gelatinase activities of MMP-2 and MMP-9 using gelatin zymography (one representative zymography of three independent experiments). (c) EMMPRIN, uPA, MMP-2, MMP-9 and TIMP-1 transcripts were quantified using quantitative RT-PCR in DOK and SCC-9 cells incubated for 24 h with CHO Mb or CHO-Emp Mb (20 μg/mL). Columns are means of gene expression relative to TBP housekeeping gene of at least three independent experiments carried out in triplicate; bars , SD. * denotes significant difference with p < 0.05. (d) SCC-9 and DOK cells were grown in microscope slide chambers to 50-60% confluence and incubated with CHO Mb or CHO-Emp Mb (20 μg/mL). After 24 h, cells were immunostained for uPA using a goat anti-human antibody (green) and counterstained with DAPI (blue). A more intense staining was observed in CHO-Emp mb treated DOK and SCC-9 cells compared to the control CHO mb treated cells. B , SCC-9 and DOK cells were incubated with 20 μg/mL of an anti-EMMPRIN blocking antibody (Ancell, Bayport, MN) or with a non-immune IgG antibody in serum-free medium. After 24 h incubation, conditioned medium was harvested for gelatinase and uPA casein zymography. Columns represent average values of the densitometric quantification from at least three independent experiments carried out in triplicate; bars , SD . * denotes significant difference with p < 0.05. C , DOK and SCC-9 cells were transfected with EMMPRIN siRNA (Emp-siRNA) or scrambled control siRNA (Ctl siRNA) at 33 nmol/L concentration. EMMPRIN and uPA mRNA expression was evaluated by quantitative RT-PCR analyses. Columns represent mean ± SD of relative expression to TBP housekeeping gene of at least 3 independent experiments carried out in triplicate; bars , SD. * denotes significant difference with p < 0.05.

Article Snippet: For immunohistochemistry, FFPE sections were stained with a mouse anti-human EMMPRIN antibody (BD-Pharmingen) and a goat anti human uPA antibody (American diagnostica).

Techniques: Transfection, Isolation, Centrifugation, Western Blot, Incubation, Activity Assay, Zymography, Quantitative RT-PCR, Expressing, Microscopy, Staining, Blocking Assay, Concentration Assay

Journal: BMC Cancer

Article Title: EMMPRIN/CD147 up-regulates urokinase-type plasminogen activator: implications in oral tumor progression

doi: 10.1186/1471-2407-12-115

Figure Lengend Snippet: EMMPRIN regulates invasion in oral dysplastic DOK and tumor SCC-9 cell lines in an MMP and uPA dependent manner. A , The in vitro invasive property of DOK and SCC-9 cells incubated with CHO-Emp membranes and treated or not with an anti-EMMPRIN blocking antibody (20 μg/mL) were compared using tissue culture Transwell inserts (8-mm pore size; BD Biosciences) placed in a 24-well culture plate. Cells incubated with CHO control or treated with an anti-IgG antibody were used as control. Cells (1 × 10 5 ) suspended in serum-free media were seeded into the upper well of each insert onto membranes coated with growth factor-reduced Matrigel (BD Biosciences). After 48 h incubation, cells that remained in the top compartment were removed by cotton swabs, and cells on the underside of insert filters were fixed, stained, and counted under a microscope. The columns shown represent average values from at least three independent experiments carried out in triplicate; bars , SD. * denotes significant difference with p < 0.05. B , DOK and SCC-9 cells were transfected with EMMPRIN siRNA or scrambled siRNA (Ctl siRNA) prior to invasion assays. The columns represent means of three independent experiments carried out in triplicate; bars , SD. C , Invasive Indices. DOK and SCC-9 cells (1 × 10 5 ) seeded into the upper well of tissue culture Transwell inserts coated with growth factor-reduced Matrigel (for invasion assay) or not coated (for migration assay) were incubated with CHO or CHO-Emp membranes (Control and CHO Emp mb respectively). After 48 h of incubation, migrating or invading cells on the underside of insert filters were fixed, stained, and counted under a microscope. Invasion Indices were calculated as percent of the invaded cells relative to the migrated cells. The columns shown represent average values from at least three independent experiments carried out in triplicate; bars , SD. * denotes significant difference with p < 0.05. D , Relative contribution of uPA and MMPs to the in vitro invasive property of DOK and SCC-9 cells. Cells (1 × 10 5 ) were seeded into the upper well of matrigel coated inserts and incubated with CHO or CHO-Emp membranes. uPA inhibitor amiloride (20 nmol/L) or MMP inhibitor marimastat (10 μmol/L) were added alone or in combination together with the membranes. After 48 h invading cells were fixed, stained, and counted. Columns represent average values from at least three independent experiments carried out in triplicate; bars , SD. * denotes significant difference with p < 0.05.

Article Snippet: For immunohistochemistry, FFPE sections were stained with a mouse anti-human EMMPRIN antibody (BD-Pharmingen) and a goat anti human uPA antibody (American diagnostica).

Techniques: In Vitro, Incubation, Blocking Assay, Staining, Microscopy, Transfection, Invasion Assay, Migration