Structured Review

Santa Cruz Biotechnology anti actin polyclonal ab i 19
Anti Actin Polyclonal Ab I 19, supplied by Santa Cruz Biotechnology, 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/anti actin polyclonal ab i 19/product/Santa Cruz Biotechnology
Average 85 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti actin polyclonal ab i 19 - by Bioz Stars, 2020-09
85/100 stars

Images

Related Articles

other:

Article Title: Expression of Prion Protein in Mouse Erythroid Progenitors and Differentiating Murine Erythroleukemia Cells
Article Snippet: The anti-actin polyclonal Ab I-19 (Santa Cruz, Santa Cruz, CA, USA) (0.5 µg/mL) was used as a loading control.

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 91
    Santa Cruz Biotechnology goat polyclonal anti actin
    HT-1080 cells containing the infectious HTLV-1 ACH.p30 II mutant provirus, defective for p30 II production, exhibit increased genomic and mitochondrial DNA-damage as compared to the wildtype ACH provirus. (A and B) The parental HT-1080 cell-line and the HT1080/HTLV-1 ACH.wt and ACH.p30 II mutant proviral clones were stained using a Click-iT Alexa Fluor 594-TUNEL kit (Molecular Probes; red signal) and a rabbit <t>polyclonal</t> primary antibody that recognizes the Ser139-phosphorylated histone variant H2A.X (Anti-Phospho-Ser139-H2A.X; green signal) which localizes at sites of genomic DNA-damage. The chemical uncoupler CCCP and DNAse I were included as positive controls. The cell nuclei were stained with the fluorescent dye, Hoechst 33342 (Molecular Probes; blue signal). The arrows in the enlarged inset panels at right indicate Phospho-Ser139-H2A.X foci in the merged images. The data in B represent the mean ± standard deviation (error bars) from three independent experiments. (C and D) HT-1080 cells and the HT-1080/HTLV-1 ACH.wt and ACH.p30 II mutant proviral clones were stained using a fluorescent TUNEL kit (red signal) and a rabbit polyclonal Anti-TOM20 primary antibody (green signal) to detect mitochondrial DNA-damage resulting from oxidative stress. The cell nuclei were stained with Hoechst 33342. Positive control samples were treated with either CCCP or DNAse I. The arrows in the enlarged inset panels at right indicate sites of mitochondrial DNA-damage. The graph in C shows the relative numbers of TUNEL/TOM20-positive foci per cell; and the data represent the mean ± standard deviation (error bars) from three independent experiments. (E and F) The ability of the HTLV-1 p30 II protein to protect against Tax/HBZ-induced cytotoxicity was determined by either (E) transfecting HT-1080 cells with Tax or HBZ expression constructs and then transducing them with lentiviral-HTLV-1 p30 II (HA) particles or an empty lentiviral vector, or (F) cotransfecting the cells with expression constructs for Tax, HBZ, and/or p30 II (HA). Certain samples were repeatedly cotransfected with a siRNA- tigar oligonucleotide or scrRNA control using HiPerFect transfection reagent. The cells in E were also treated with staurosporine (100 nM) as a positive control to induce apoptosis. Apoptotic cells were detected by staining them with Annexin V-FITC (green signal) and propidium iodide (PI; red signal); and the relative percentages of Annexin V-FITC +/− PI-positive cells were quantified by fluorescence-microscopy, as compared to the total numbers of cells visualized using a DIC phase-contrast filter. All the data is representative of at least three independent experiments; and the data in F represent the mean of the experiments ± standard deviation (error bars).
    Goat Polyclonal Anti Actin, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat polyclonal anti actin/product/Santa Cruz Biotechnology
    Average 91 stars, based on 45 article reviews
    Price from $9.99 to $1999.99
    goat polyclonal anti actin - by Bioz Stars, 2020-09
    91/100 stars
      Buy from Supplier

    91
    Santa Cruz Biotechnology rabbit polyclonal anti actin
    Co-localization and co-immunoprecipitation of cyclophilin B with NaDC1. A , subcellular localization of CypB and NaDC1 in non-transfected HEK293 cells treated ± 200 ng/ml brefeldin A , Western analyses of the last seven fractions out of 10 (numbered 1–7), in which GM130 ( cis -Golgi Matrix protein 130; cis -Golgi marker) and PDI (ER marker) were immunodetected. In non-treated cells, CypB co-localized with immature NaDC1 in the ER (fractions 6 and 7). In brefeldin A-treated cells, the expression of the immature NaDC1 form and CypB was increased in the ER (fractions 6 and 7). B , co-immunoprecipitation of CypB with NaDC1: HA-tagged CypB was co-injected with either FLAG-tagged NaDC1 or wt TRPV6 (positive control). Non-injected oocytes were used as negative control. NaDC1 and TRPV6 were immunoprecipitated (IP) with rabbit <t>polyclonal</t> anti-FLAG and chicken polyclonal anti-huTRPV6 antibodies, respectively. Western membranes were immunoblotted (IB) with the mouse monoclononal anti-HA. A CypB immunoreactive band (∼20 kDa) was detected in all co-immunoprecipitation studies.
    Rabbit Polyclonal Anti Actin, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 41 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti actin/product/Santa Cruz Biotechnology
    Average 91 stars, based on 41 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal anti actin - by Bioz Stars, 2020-09
    91/100 stars
      Buy from Supplier

    Image Search Results


    HT-1080 cells containing the infectious HTLV-1 ACH.p30 II mutant provirus, defective for p30 II production, exhibit increased genomic and mitochondrial DNA-damage as compared to the wildtype ACH provirus. (A and B) The parental HT-1080 cell-line and the HT1080/HTLV-1 ACH.wt and ACH.p30 II mutant proviral clones were stained using a Click-iT Alexa Fluor 594-TUNEL kit (Molecular Probes; red signal) and a rabbit polyclonal primary antibody that recognizes the Ser139-phosphorylated histone variant H2A.X (Anti-Phospho-Ser139-H2A.X; green signal) which localizes at sites of genomic DNA-damage. The chemical uncoupler CCCP and DNAse I were included as positive controls. The cell nuclei were stained with the fluorescent dye, Hoechst 33342 (Molecular Probes; blue signal). The arrows in the enlarged inset panels at right indicate Phospho-Ser139-H2A.X foci in the merged images. The data in B represent the mean ± standard deviation (error bars) from three independent experiments. (C and D) HT-1080 cells and the HT-1080/HTLV-1 ACH.wt and ACH.p30 II mutant proviral clones were stained using a fluorescent TUNEL kit (red signal) and a rabbit polyclonal Anti-TOM20 primary antibody (green signal) to detect mitochondrial DNA-damage resulting from oxidative stress. The cell nuclei were stained with Hoechst 33342. Positive control samples were treated with either CCCP or DNAse I. The arrows in the enlarged inset panels at right indicate sites of mitochondrial DNA-damage. The graph in C shows the relative numbers of TUNEL/TOM20-positive foci per cell; and the data represent the mean ± standard deviation (error bars) from three independent experiments. (E and F) The ability of the HTLV-1 p30 II protein to protect against Tax/HBZ-induced cytotoxicity was determined by either (E) transfecting HT-1080 cells with Tax or HBZ expression constructs and then transducing them with lentiviral-HTLV-1 p30 II (HA) particles or an empty lentiviral vector, or (F) cotransfecting the cells with expression constructs for Tax, HBZ, and/or p30 II (HA). Certain samples were repeatedly cotransfected with a siRNA- tigar oligonucleotide or scrRNA control using HiPerFect transfection reagent. The cells in E were also treated with staurosporine (100 nM) as a positive control to induce apoptosis. Apoptotic cells were detected by staining them with Annexin V-FITC (green signal) and propidium iodide (PI; red signal); and the relative percentages of Annexin V-FITC +/− PI-positive cells were quantified by fluorescence-microscopy, as compared to the total numbers of cells visualized using a DIC phase-contrast filter. All the data is representative of at least three independent experiments; and the data in F represent the mean of the experiments ± standard deviation (error bars).

    Journal: Virology

    Article Title: The TP53-Induced Glycolysis and Apoptosis Regulator mediates cooperation between HTLV-1 p30II and the retroviral oncoproteins Tax and HBZ and is highly expressed in an in vivo xenograft model of HTLV-1-induced lymphoma

    doi: 10.1016/j.virol.2018.05.007

    Figure Lengend Snippet: HT-1080 cells containing the infectious HTLV-1 ACH.p30 II mutant provirus, defective for p30 II production, exhibit increased genomic and mitochondrial DNA-damage as compared to the wildtype ACH provirus. (A and B) The parental HT-1080 cell-line and the HT1080/HTLV-1 ACH.wt and ACH.p30 II mutant proviral clones were stained using a Click-iT Alexa Fluor 594-TUNEL kit (Molecular Probes; red signal) and a rabbit polyclonal primary antibody that recognizes the Ser139-phosphorylated histone variant H2A.X (Anti-Phospho-Ser139-H2A.X; green signal) which localizes at sites of genomic DNA-damage. The chemical uncoupler CCCP and DNAse I were included as positive controls. The cell nuclei were stained with the fluorescent dye, Hoechst 33342 (Molecular Probes; blue signal). The arrows in the enlarged inset panels at right indicate Phospho-Ser139-H2A.X foci in the merged images. The data in B represent the mean ± standard deviation (error bars) from three independent experiments. (C and D) HT-1080 cells and the HT-1080/HTLV-1 ACH.wt and ACH.p30 II mutant proviral clones were stained using a fluorescent TUNEL kit (red signal) and a rabbit polyclonal Anti-TOM20 primary antibody (green signal) to detect mitochondrial DNA-damage resulting from oxidative stress. The cell nuclei were stained with Hoechst 33342. Positive control samples were treated with either CCCP or DNAse I. The arrows in the enlarged inset panels at right indicate sites of mitochondrial DNA-damage. The graph in C shows the relative numbers of TUNEL/TOM20-positive foci per cell; and the data represent the mean ± standard deviation (error bars) from three independent experiments. (E and F) The ability of the HTLV-1 p30 II protein to protect against Tax/HBZ-induced cytotoxicity was determined by either (E) transfecting HT-1080 cells with Tax or HBZ expression constructs and then transducing them with lentiviral-HTLV-1 p30 II (HA) particles or an empty lentiviral vector, or (F) cotransfecting the cells with expression constructs for Tax, HBZ, and/or p30 II (HA). Certain samples were repeatedly cotransfected with a siRNA- tigar oligonucleotide or scrRNA control using HiPerFect transfection reagent. The cells in E were also treated with staurosporine (100 nM) as a positive control to induce apoptosis. Apoptotic cells were detected by staining them with Annexin V-FITC (green signal) and propidium iodide (PI; red signal); and the relative percentages of Annexin V-FITC +/− PI-positive cells were quantified by fluorescence-microscopy, as compared to the total numbers of cells visualized using a DIC phase-contrast filter. All the data is representative of at least three independent experiments; and the data in F represent the mean of the experiments ± standard deviation (error bars).

    Article Snippet: The following primary antibodies were used throughout: goat polyclonal Anti-HTLV-1 Tax (vC-12; Santa Cruz Biotechnology), goat polyclonal Anti-Actin (I-19; Santa Cruz Biotechnology), mouse monoclonal Anti-Beclin-1 (E-8; Santa Cruz Biotechnology), goat polyclonal Anti-TIM23 (C-19; Santa Cruz Biotechnology), rabbit polyclonal Anti-TOM20 (FL-145; Santa Cruz Biotechnology), mouse monoclonal Anti-SQSTM1 (D-3; Santa Cruz Biotechnology), rabbit polyclonal Anti-Phospho-Ser139-H2A.X (Santa Cruz Biotechnology), rabbit polyclonal Anti-GFP (FL; Santa Cruz Biotechnology), Anti-HTLV-1 p19Gag (Zeptometrix), mouse monoclonal Anti-TIGAR (G-2; Santa Cruz Biotechnology), rabbit polyclonal Anti-Human TIGAR (described in ), rabbit polyclonal Anti-Human Ki67 (huKi67, H-300; Santa Cruz Biotechnology), mouse monoclonal Anti-FLAG M2 (Sigma-Aldrich), mouse monoclonal Anti-c-Myc (9E10; Santa Cruz Biotechnology), mouse monoclonal Anti-CD31/PECAM1 (H-3; Santa Cruz Biotechnology), mouse monoclonal Anti-Flk1/VEGFR2 (A-3; Santa Cruz Biotechnology), mouse monoclonal Anti-HIF-1α (28b; Santa Cruz Biotechnology), and mouse monoclonal Anti-VEGF (C-1; Santa Cruz Biotechnology).

    Techniques: Mutagenesis, Clone Assay, Staining, TUNEL Assay, Variant Assay, Standard Deviation, Positive Control, Expressing, Construct, Plasmid Preparation, Transfection, Fluorescence, Microscopy

    Figure 2. PrP populations with and without the GPI-anchor are shed into the cell medium from PrP expressing cells in addition to the N1 fragment. Multiplex western immunoblot detection of cell medium from PrPwt (lanes 2, 4 and 6) and PrPΔ121–123 (lanes 3, 5 and 7) expressing cells. The samples were treated with PNGase F to remove N-linked oligosaccharides. The PVDF membrane was probed with both the N-terminal polyclonal rabbit antibody R505 and the C-terminal monoclonal mouse antibody L42, followed by IRDye 800 goat anti-rabbit (green) and Alexa Fluor 680 goat anti-mouse (red). Both colors were imaged in a single scan. The N-terminal antibody (green, lanes 2 and 3) detected full-length PrP with intact GPI-anchor (FL), GPI-anchorless full-length (FL-S), and the N-terminal fragment (N1) and the C-terminal antibody (red, lanes 4 and 5) detected the FL, FL-S, the truncated C-terminal fragments with intact GPI-anchor (C1) and without GPI-anchor (C1-S). Yellow indicates merged overlapping colors (lanes 6 and 7). Approximate molecular masses are indicated on the left-hand side (kDa).

    Journal: Prion

    Article Title: Separate mechanisms act concurrently to shed and release the prion protein from the cell

    doi: 10.4161/pri.22588

    Figure Lengend Snippet: Figure 2. PrP populations with and without the GPI-anchor are shed into the cell medium from PrP expressing cells in addition to the N1 fragment. Multiplex western immunoblot detection of cell medium from PrPwt (lanes 2, 4 and 6) and PrPΔ121–123 (lanes 3, 5 and 7) expressing cells. The samples were treated with PNGase F to remove N-linked oligosaccharides. The PVDF membrane was probed with both the N-terminal polyclonal rabbit antibody R505 and the C-terminal monoclonal mouse antibody L42, followed by IRDye 800 goat anti-rabbit (green) and Alexa Fluor 680 goat anti-mouse (red). Both colors were imaged in a single scan. The N-terminal antibody (green, lanes 2 and 3) detected full-length PrP with intact GPI-anchor (FL), GPI-anchorless full-length (FL-S), and the N-terminal fragment (N1) and the C-terminal antibody (red, lanes 4 and 5) detected the FL, FL-S, the truncated C-terminal fragments with intact GPI-anchor (C1) and without GPI-anchor (C1-S). Yellow indicates merged overlapping colors (lanes 6 and 7). Approximate molecular masses are indicated on the left-hand side (kDa).

    Article Snippet: Goat polyclonal anti-Actin antiserum (I-19; Santa Cruz Biotechnology) is raised against a peptide mapping at the C-terminus of actin of human origin.

    Techniques: Expressing, Multiplex Assay, Western Blot

    p120-uncoupled E-cadherin is less able to suppress invasiveness. (A) Ectopic E-cadherin recruits p120 to cell junctions. MDA-231 cells were infected with retroviruses expressing neomycin resistance alone or in combination with wt E-cadherin. More than 95% of G418-selected cells expressed wt E-cadherin. Wt E-cadherin accumulated at areas of cell–cell contact (mAb HECD-1). Endogenous p120 was primarily cytoplasmic in control cells and was recruited to cell–cell junctions in cells expressing wt E-cadherin (detected with polyclonal antibody F1αSH). (B) E-cadherin expression suppresses cell migration. After overnight serum starvation, confluent monolayers of control neo cells or MDA-231 cells expressing wt E-cadherin were scratched with a pipette tip, and cell migration into the wound was monitored over time in the presence of HGF. The number of individual cells that crossed the original cell front after 12 h was counted, and results were expressed as a percentage of migrating cells compared with neo control ( n = 6). (C) E-cadherin suppresses cell invasiveness. The invasiveness of E-cadherin–expressing MDA-231 or UMRC3 cells was tested in vitro using HGF (MDA-231) or serum (UMRC3) as the chemoattractant ( n = 6). (D) p120-uncoupled E-cadherin is less able to suppress invasiveness. Polyclonal populations of MDA-231 cells expressing neo control, wt E-cadherin, or a p120-uncoupled E-cadherin mutant (764-AAA) were subjected to invasion assays in response to HGF ( n = 6). Error bars indicate SEM. **, P

    Journal: The Journal of Cell Biology

    Article Title: p120 catenin is essential for mesenchymal cadherin-mediated regulation of cell motility and invasiveness

    doi: 10.1083/jcb.200605022

    Figure Lengend Snippet: p120-uncoupled E-cadherin is less able to suppress invasiveness. (A) Ectopic E-cadherin recruits p120 to cell junctions. MDA-231 cells were infected with retroviruses expressing neomycin resistance alone or in combination with wt E-cadherin. More than 95% of G418-selected cells expressed wt E-cadherin. Wt E-cadherin accumulated at areas of cell–cell contact (mAb HECD-1). Endogenous p120 was primarily cytoplasmic in control cells and was recruited to cell–cell junctions in cells expressing wt E-cadherin (detected with polyclonal antibody F1αSH). (B) E-cadherin expression suppresses cell migration. After overnight serum starvation, confluent monolayers of control neo cells or MDA-231 cells expressing wt E-cadherin were scratched with a pipette tip, and cell migration into the wound was monitored over time in the presence of HGF. The number of individual cells that crossed the original cell front after 12 h was counted, and results were expressed as a percentage of migrating cells compared with neo control ( n = 6). (C) E-cadherin suppresses cell invasiveness. The invasiveness of E-cadherin–expressing MDA-231 or UMRC3 cells was tested in vitro using HGF (MDA-231) or serum (UMRC3) as the chemoattractant ( n = 6). (D) p120-uncoupled E-cadherin is less able to suppress invasiveness. Polyclonal populations of MDA-231 cells expressing neo control, wt E-cadherin, or a p120-uncoupled E-cadherin mutant (764-AAA) were subjected to invasion assays in response to HGF ( n = 6). Error bars indicate SEM. **, P

    Article Snippet: Primary antibodies were used as follows: 0.25 μg/ml anti-p120 mAbs pp120 and 1 μg/ml 8D11 (does not recognize human p120), anti–E-cadherin mAbs (1/2,500; C-20820; BD Biosciences) and 1 μg/ml HECD-1 (Zymed Laboratories), 1 μg/ml anti–c-Met (C-28; Santa Cruz Biotechnology, Inc.), anti–β-catenin polyclonal antibody (1/1000; C2206; Sigma-Aldrich), 5 μg/ml anti-Flag tag mAb (M2; Sigma-Aldrich), 1 μg/ml anti-myc tag (9E10; Sigma-Aldrich), anti–cadherin 11 antibodies (WTID1 [polyclonal antibody] and 5B2H5 [mAb]; Zymed Laboratories), and 0.6 μg/ml anti-actin goat polyclonal antibody (I-19; Santa Cruz Biotechnology, Inc.).

    Techniques: Multiple Displacement Amplification, Infection, Expressing, Migration, Transferring, In Vitro, Mutagenesis

    A cadherin-uncoupled p120 mutant does not promote motility and invasiveness. (A) Ectopic expression of murine p120 rescues the migration of p120-depleted cells. p120-depleted MDA-231 cells (clone 3; Fig. 1 B ) were infected with control neo retrovirus or retrovirus expressing murine p120. After G418 selection, stable polyclonal cell lines were used in a scratch-wound assay to test their migration in response to HGF. Images shown are 0 and 12 h after HGF addition. The line denotes the cell front at time 0. The number of individual cells that crossed the line after 12 h was counted, and results were expressed as a percentage of migrating cells compared with control (MDA-231-pRS). n = 9. (B) The increased migration of p120-expressing cells is not due to reduced cell–cell adhesion. Cells were suspended as hanging drops and allowed to aggregate overnight. The strength of cell adhesion was assessed after passing the cell aggregates 10 times through a standard 200-μl pipette tip. Note that p120-expressing cells are more resistant to dissaggregation than cells depleted of endogenous p120. (C) MDA-231 cells with knocked down expression of endogenous p120 (clone 3) were infected with retroviruses expressing neo resistance alone or together with full-length murine p120 or a murine p120 mutant (mp120-A1), which is unable to bind E-cadherin. The invasiveness of p120-reexpressing cells was tested in vitro toward HGF ( n = 6). (bottom) Lysates from all cell lines were subjected to SDS-PAGE and Western blotted for expression of p120 (using mAb pp120) and actin. Error bars indicate SEM. **, P

    Journal: The Journal of Cell Biology

    Article Title: p120 catenin is essential for mesenchymal cadherin-mediated regulation of cell motility and invasiveness

    doi: 10.1083/jcb.200605022

    Figure Lengend Snippet: A cadherin-uncoupled p120 mutant does not promote motility and invasiveness. (A) Ectopic expression of murine p120 rescues the migration of p120-depleted cells. p120-depleted MDA-231 cells (clone 3; Fig. 1 B ) were infected with control neo retrovirus or retrovirus expressing murine p120. After G418 selection, stable polyclonal cell lines were used in a scratch-wound assay to test their migration in response to HGF. Images shown are 0 and 12 h after HGF addition. The line denotes the cell front at time 0. The number of individual cells that crossed the line after 12 h was counted, and results were expressed as a percentage of migrating cells compared with control (MDA-231-pRS). n = 9. (B) The increased migration of p120-expressing cells is not due to reduced cell–cell adhesion. Cells were suspended as hanging drops and allowed to aggregate overnight. The strength of cell adhesion was assessed after passing the cell aggregates 10 times through a standard 200-μl pipette tip. Note that p120-expressing cells are more resistant to dissaggregation than cells depleted of endogenous p120. (C) MDA-231 cells with knocked down expression of endogenous p120 (clone 3) were infected with retroviruses expressing neo resistance alone or together with full-length murine p120 or a murine p120 mutant (mp120-A1), which is unable to bind E-cadherin. The invasiveness of p120-reexpressing cells was tested in vitro toward HGF ( n = 6). (bottom) Lysates from all cell lines were subjected to SDS-PAGE and Western blotted for expression of p120 (using mAb pp120) and actin. Error bars indicate SEM. **, P

    Article Snippet: Primary antibodies were used as follows: 0.25 μg/ml anti-p120 mAbs pp120 and 1 μg/ml 8D11 (does not recognize human p120), anti–E-cadherin mAbs (1/2,500; C-20820; BD Biosciences) and 1 μg/ml HECD-1 (Zymed Laboratories), 1 μg/ml anti–c-Met (C-28; Santa Cruz Biotechnology, Inc.), anti–β-catenin polyclonal antibody (1/1000; C2206; Sigma-Aldrich), 5 μg/ml anti-Flag tag mAb (M2; Sigma-Aldrich), 1 μg/ml anti-myc tag (9E10; Sigma-Aldrich), anti–cadherin 11 antibodies (WTID1 [polyclonal antibody] and 5B2H5 [mAb]; Zymed Laboratories), and 0.6 μg/ml anti-actin goat polyclonal antibody (I-19; Santa Cruz Biotechnology, Inc.).

    Techniques: Mutagenesis, Expressing, Migration, Multiple Displacement Amplification, Infection, Selection, Scratch Wound Assay Assay, Transferring, In Vitro, SDS Page, Western Blot

    Endogenous mesenchymal cadherins promote cell invasiveness. (A) Expression of the E-cadherin juxtamembrane domain inhibits cadherin 11 levels and blocks invasiveness. MDA-231 cells were infected with control retrovirus (zeo) or retrovirus expressing a small, p120-binding, myc-tagged fragment of the E-cadherin cytoplasmic tail (ΔCB) that cannot associate with β-catenin. Polyclonal stable cell lines were subjected to invasion assays, toward a gradient of HGF ( n = 6). (bottom) Lysates from both cell lines were subjected to SDS-PAGE and Western blotted for expression of cadherin 11, ΔCB (using the myc tag–specific mAb 9E10), and actin. (B) p120 regulates the levels of endogenous mesenchymal cadherins. Polyclonal populations of p120-depleted MDA-231 or UMRC3 cells were examined for their expression of cadherin 11 or N-cadherin, respectively. Note that upon p120 depletion (top), levels of endogenous mesenchymal cadherins (middle and graphs) are reduced ( n = 3). (C) Endogenous mesenchymal cadherins promote cell invasiveness. MDA-231 and UMRC3 cells were transiently transfected by electroporation with either control siRNA or siRNA specific for human cadherin 11 or N-cadherin, respectively. Control experiments verified maximal knock down of cadherin expression 3 d after transfection. 2 d after transfection, cells were serum starved overnight and plated in Matrigel-coated transwells. Invasiveness was determined 24 h later in response to a gradient of HGF (MDA-231) or serum (UMRC3). n = 6. Error bars indicate SEM. *, P

    Journal: The Journal of Cell Biology

    Article Title: p120 catenin is essential for mesenchymal cadherin-mediated regulation of cell motility and invasiveness

    doi: 10.1083/jcb.200605022

    Figure Lengend Snippet: Endogenous mesenchymal cadherins promote cell invasiveness. (A) Expression of the E-cadherin juxtamembrane domain inhibits cadherin 11 levels and blocks invasiveness. MDA-231 cells were infected with control retrovirus (zeo) or retrovirus expressing a small, p120-binding, myc-tagged fragment of the E-cadherin cytoplasmic tail (ΔCB) that cannot associate with β-catenin. Polyclonal stable cell lines were subjected to invasion assays, toward a gradient of HGF ( n = 6). (bottom) Lysates from both cell lines were subjected to SDS-PAGE and Western blotted for expression of cadherin 11, ΔCB (using the myc tag–specific mAb 9E10), and actin. (B) p120 regulates the levels of endogenous mesenchymal cadherins. Polyclonal populations of p120-depleted MDA-231 or UMRC3 cells were examined for their expression of cadherin 11 or N-cadherin, respectively. Note that upon p120 depletion (top), levels of endogenous mesenchymal cadherins (middle and graphs) are reduced ( n = 3). (C) Endogenous mesenchymal cadherins promote cell invasiveness. MDA-231 and UMRC3 cells were transiently transfected by electroporation with either control siRNA or siRNA specific for human cadherin 11 or N-cadherin, respectively. Control experiments verified maximal knock down of cadherin expression 3 d after transfection. 2 d after transfection, cells were serum starved overnight and plated in Matrigel-coated transwells. Invasiveness was determined 24 h later in response to a gradient of HGF (MDA-231) or serum (UMRC3). n = 6. Error bars indicate SEM. *, P

    Article Snippet: Primary antibodies were used as follows: 0.25 μg/ml anti-p120 mAbs pp120 and 1 μg/ml 8D11 (does not recognize human p120), anti–E-cadherin mAbs (1/2,500; C-20820; BD Biosciences) and 1 μg/ml HECD-1 (Zymed Laboratories), 1 μg/ml anti–c-Met (C-28; Santa Cruz Biotechnology, Inc.), anti–β-catenin polyclonal antibody (1/1000; C2206; Sigma-Aldrich), 5 μg/ml anti-Flag tag mAb (M2; Sigma-Aldrich), 1 μg/ml anti-myc tag (9E10; Sigma-Aldrich), anti–cadherin 11 antibodies (WTID1 [polyclonal antibody] and 5B2H5 [mAb]; Zymed Laboratories), and 0.6 μg/ml anti-actin goat polyclonal antibody (I-19; Santa Cruz Biotechnology, Inc.).

    Techniques: Expressing, Multiple Displacement Amplification, Infection, Binding Assay, Stable Transfection, SDS Page, Western Blot, Transfection, Electroporation

    Endogenous p120 promotes invasiveness. (A) p120 depletion blocks invasiveness. E-cadherin–deficient MDA-231 and UMRC3 cells were infected with control pRS retrovirus or with a retrovirus expressing anti-human p120-specific shRNA. Polyclonal cell populations were generated, and invasion assays were performed in vitro using 20 ng/ml HGF as a chemoattractant for MDA-231 cells or 5% FBS for the UMRC3 cells. After a 20-h incubation, cells on the underside of a Matrigel-coated transwell membrane were counted under a 20× objective ( n = 6). (B) Endogenous p120 levels are directly correlated to cell invasion. MDA-231 cells were infected with control retrovirus or retrovirus expressing p120-specific shRNA. Individual clones of shRNA-expressing cells were selected and tested for levels of endogenous p120 and invasiveness in vitro, toward HGF ( n = 6). (bottom) Lysates from all cell lines were subjected to SDS-PAGE and Western blotted for expression of endogenous p120 (using mAb pp120) and actin. The two p120 bands correspond to p120 isoforms 1 (top) and 3 (bottom). Error bars indicate SEM. *, P

    Journal: The Journal of Cell Biology

    Article Title: p120 catenin is essential for mesenchymal cadherin-mediated regulation of cell motility and invasiveness

    doi: 10.1083/jcb.200605022

    Figure Lengend Snippet: Endogenous p120 promotes invasiveness. (A) p120 depletion blocks invasiveness. E-cadherin–deficient MDA-231 and UMRC3 cells were infected with control pRS retrovirus or with a retrovirus expressing anti-human p120-specific shRNA. Polyclonal cell populations were generated, and invasion assays were performed in vitro using 20 ng/ml HGF as a chemoattractant for MDA-231 cells or 5% FBS for the UMRC3 cells. After a 20-h incubation, cells on the underside of a Matrigel-coated transwell membrane were counted under a 20× objective ( n = 6). (B) Endogenous p120 levels are directly correlated to cell invasion. MDA-231 cells were infected with control retrovirus or retrovirus expressing p120-specific shRNA. Individual clones of shRNA-expressing cells were selected and tested for levels of endogenous p120 and invasiveness in vitro, toward HGF ( n = 6). (bottom) Lysates from all cell lines were subjected to SDS-PAGE and Western blotted for expression of endogenous p120 (using mAb pp120) and actin. The two p120 bands correspond to p120 isoforms 1 (top) and 3 (bottom). Error bars indicate SEM. *, P

    Article Snippet: Primary antibodies were used as follows: 0.25 μg/ml anti-p120 mAbs pp120 and 1 μg/ml 8D11 (does not recognize human p120), anti–E-cadherin mAbs (1/2,500; C-20820; BD Biosciences) and 1 μg/ml HECD-1 (Zymed Laboratories), 1 μg/ml anti–c-Met (C-28; Santa Cruz Biotechnology, Inc.), anti–β-catenin polyclonal antibody (1/1000; C2206; Sigma-Aldrich), 5 μg/ml anti-Flag tag mAb (M2; Sigma-Aldrich), 1 μg/ml anti-myc tag (9E10; Sigma-Aldrich), anti–cadherin 11 antibodies (WTID1 [polyclonal antibody] and 5B2H5 [mAb]; Zymed Laboratories), and 0.6 μg/ml anti-actin goat polyclonal antibody (I-19; Santa Cruz Biotechnology, Inc.).

    Techniques: Multiple Displacement Amplification, Infection, Expressing, shRNA, Generated, In Vitro, Incubation, Clone Assay, SDS Page, Western Blot

    Co-localization and co-immunoprecipitation of cyclophilin B with NaDC1. A , subcellular localization of CypB and NaDC1 in non-transfected HEK293 cells treated ± 200 ng/ml brefeldin A , Western analyses of the last seven fractions out of 10 (numbered 1–7), in which GM130 ( cis -Golgi Matrix protein 130; cis -Golgi marker) and PDI (ER marker) were immunodetected. In non-treated cells, CypB co-localized with immature NaDC1 in the ER (fractions 6 and 7). In brefeldin A-treated cells, the expression of the immature NaDC1 form and CypB was increased in the ER (fractions 6 and 7). B , co-immunoprecipitation of CypB with NaDC1: HA-tagged CypB was co-injected with either FLAG-tagged NaDC1 or wt TRPV6 (positive control). Non-injected oocytes were used as negative control. NaDC1 and TRPV6 were immunoprecipitated (IP) with rabbit polyclonal anti-FLAG and chicken polyclonal anti-huTRPV6 antibodies, respectively. Western membranes were immunoblotted (IB) with the mouse monoclononal anti-HA. A CypB immunoreactive band (∼20 kDa) was detected in all co-immunoprecipitation studies.

    Journal: The Journal of Biological Chemistry

    Article Title: Synthesis, Maturation, and Trafficking of Human Na+-Dicarboxylate Cotransporter NaDC1 Requires the Chaperone Activity of Cyclophilin B *

    doi: 10.1074/jbc.M110.171728

    Figure Lengend Snippet: Co-localization and co-immunoprecipitation of cyclophilin B with NaDC1. A , subcellular localization of CypB and NaDC1 in non-transfected HEK293 cells treated ± 200 ng/ml brefeldin A , Western analyses of the last seven fractions out of 10 (numbered 1–7), in which GM130 ( cis -Golgi Matrix protein 130; cis -Golgi marker) and PDI (ER marker) were immunodetected. In non-treated cells, CypB co-localized with immature NaDC1 in the ER (fractions 6 and 7). In brefeldin A-treated cells, the expression of the immature NaDC1 form and CypB was increased in the ER (fractions 6 and 7). B , co-immunoprecipitation of CypB with NaDC1: HA-tagged CypB was co-injected with either FLAG-tagged NaDC1 or wt TRPV6 (positive control). Non-injected oocytes were used as negative control. NaDC1 and TRPV6 were immunoprecipitated (IP) with rabbit polyclonal anti-FLAG and chicken polyclonal anti-huTRPV6 antibodies, respectively. Western membranes were immunoblotted (IB) with the mouse monoclononal anti-HA. A CypB immunoreactive band (∼20 kDa) was detected in all co-immunoprecipitation studies.

    Article Snippet: The primary antibodies mouse polyclonal anti-SLC13A2 (Abnova), rabbit polyclonal anti-cyclophilin B (Pierce), mouse monoclonal anti-HA (Sigma) and rabbit polyclonal anti-actin (I-19; Santa Cruz Biotechnology) were used at a dilution of 1/1,000.

    Techniques: Immunoprecipitation, Transfection, Western Blot, Marker, Expressing, Injection, Positive Control, Negative Control