scf protein expression  (Millipore)


Bioz Verified Symbol Millipore is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Name:
    Protein Expression
    Description:
    Details of the expression of cloned DNA or RNA templates in all the major in vivo and in vitro systems are included such as cultured mammalian cells the yeasts Saccharomyces cerevisiae and Pichia pastoris baculovirus Xenopus oocytes and prokaryotic cells Cell free systems of both eukaryotes and prokaryotes are described in addition to the prokaryotic systems that offer coupled transcription translation
    Catalog Number:
    z340820
    Price:
    None
    Buy from Supplier


    Structured Review

    Millipore scf protein expression
    Protein Expression
    Details of the expression of cloned DNA or RNA templates in all the major in vivo and in vitro systems are included such as cultured mammalian cells the yeasts Saccharomyces cerevisiae and Pichia pastoris baculovirus Xenopus oocytes and prokaryotic cells Cell free systems of both eukaryotes and prokaryotes are described in addition to the prokaryotic systems that offer coupled transcription translation
    https://www.bioz.com/result/scf protein expression/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    scf protein expression - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Human airway epithelial cell determinants of survival and functional phenotype for primary human mast cells"

    Article Title: Human airway epithelial cell determinants of survival and functional phenotype for primary human mast cells

    Journal:

    doi: 10.1073/pnas.0503948102

    MC protease expression identified by immunostaining over time. MCs were recovered after 2 d of culture in liquid suspension culture with SCF alone, in direct coculture with HAECs, in HAEC coculture separated by a transwell membrane, with HAEC-conditioned
    Figure Legend Snippet: MC protease expression identified by immunostaining over time. MCs were recovered after 2 d of culture in liquid suspension culture with SCF alone, in direct coculture with HAECs, in HAEC coculture separated by a transwell membrane, with HAEC-conditioned

    Techniques Used: Expressing, Immunostaining

    SCF protein expression by HAECs. Release of soluble SCF protein into HAEC culture media was detected by SCF-ELISA. ( A ) All data are mean ± SEM ( n = 4). Detection of membrane-bound SCF expression by immunohistochemical staining of HAECs by using
    Figure Legend Snippet: SCF protein expression by HAECs. Release of soluble SCF protein into HAEC culture media was detected by SCF-ELISA. ( A ) All data are mean ± SEM ( n = 4). Detection of membrane-bound SCF expression by immunohistochemical staining of HAECs by using

    Techniques Used: Expressing, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Staining

    Expression of SCF mRNA by HAECs. Soluble (572 bp) and membrane-bound (488 bp) SCF expression by HAECs was identified by RT-PCR analysis. SCF cDNA was amplified by 35 cycles of PCR. Lanes 1 and 2, HAEC donor 1; lanes 3 and 4, primary human MC; lanes 5
    Figure Legend Snippet: Expression of SCF mRNA by HAECs. Soluble (572 bp) and membrane-bound (488 bp) SCF expression by HAECs was identified by RT-PCR analysis. SCF cDNA was amplified by 35 cycles of PCR. Lanes 1 and 2, HAEC donor 1; lanes 3 and 4, primary human MC; lanes 5

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Amplification, Polymerase Chain Reaction

    Related Articles

    Nucleic Acid Electrophoresis:

    Article Title: Notch1 signaling inhibits growth of EC109 esophageal carcinoma cells through downmodulation of HPV18 E6/E7 gene expression
    Article Snippet: .. Detection of p53 protein expression in EC109 cells by Western blot analysis The proteins from cell lysates of EC109 cells were separated by SDS-polyacrylamide gel electrophoresis and transferred onto PVDF membranes (Millipore, Bedford, MA, USA). .. The blots were incubated with the anti-p53 antibody or the anti-β-actin antibody (1:100 dilution in a blocking solution) at 4 °C overnight, then washed with a solution containing 5% milk, 20 mmol/L Tris-HCl (pH 7.5), 500 mmol/L NaCl, and 0.05% Tween-20 (TBS-T).

    Transfection:

    Article Title: KCa1.1, a calcium-activated potassium channel subunit alpha 1, is targeted by miR-17-5p and modulates cell migration in malignant pleural mesothelioma
    Article Snippet: .. Immunofluorescence staining of KCa1.1 protein expression Cells transfected with KCNMA1-specific siRNA, miR-17-5p mimic or control were fixed in paraformaldehyde solution (4 % v/v in PBS, Sigma, St. Louis, MO, USA) for 15 min, washed three times with PBS and permeabilized with 0.2 % Triton X-100 in PBS for 5 min. ..

    Positron Emission Tomography:

    Article Title: Ha83, a Chitin Binding Domain Encoding Gene, Is Important to Helicoverpa armigera Nucleopolyhedrovirus Budded Virus Production and Occlusion Body Assembling
    Article Snippet: .. Six truncated Ha83 protein expression vectors (pET-28a-TC151, pET-28a-TC142, pET-28a-TC128, pET-28a-TC118, pET-28a-TC113, and pET-28a-TC71) were constructed by insertion of the truncated ha83 PCR products into Bam HI and Xho I sites of the pET-28a(+) vector (Novagen, GER) with specific primers (Ha83-F used as forward primer, Ha83C151, Ha83C142, Ha83C128, Ha83C118, Ha83C113, and Ha83C71 were used as reverse primers, respectively) ( ). pET-28a-Ha83 which can encode the entire Ha83 was constructed in a similar way with primers Ha83-F and Ha83-R. And pET-28a-ha81, which contains the entire CDs of ha81 gene, was constructed with primers Ha81-F and Ha81-R as described above and used as the negative control plasmid for ha81 sharing a similar base pair number with ha83 . .. The recombinant Ha83, truncated proteins (TC151, TC142, TC128, TC118, TC113, and TC71), as well as the negative control protein Ha81 were then induced by 1 mM isopropyl-β-d-thiogalactoside (IPTG) (Sigma, USA) and purified using affinity chromatography of cOmplete His-Tag Purification Resin (Roche, SUI).

    Negative Control:

    Article Title: Ha83, a Chitin Binding Domain Encoding Gene, Is Important to Helicoverpa armigera Nucleopolyhedrovirus Budded Virus Production and Occlusion Body Assembling
    Article Snippet: .. Six truncated Ha83 protein expression vectors (pET-28a-TC151, pET-28a-TC142, pET-28a-TC128, pET-28a-TC118, pET-28a-TC113, and pET-28a-TC71) were constructed by insertion of the truncated ha83 PCR products into Bam HI and Xho I sites of the pET-28a(+) vector (Novagen, GER) with specific primers (Ha83-F used as forward primer, Ha83C151, Ha83C142, Ha83C128, Ha83C118, Ha83C113, and Ha83C71 were used as reverse primers, respectively) ( ). pET-28a-Ha83 which can encode the entire Ha83 was constructed in a similar way with primers Ha83-F and Ha83-R. And pET-28a-ha81, which contains the entire CDs of ha81 gene, was constructed with primers Ha81-F and Ha81-R as described above and used as the negative control plasmid for ha81 sharing a similar base pair number with ha83 . .. The recombinant Ha83, truncated proteins (TC151, TC142, TC128, TC118, TC113, and TC71), as well as the negative control protein Ha81 were then induced by 1 mM isopropyl-β-d-thiogalactoside (IPTG) (Sigma, USA) and purified using affinity chromatography of cOmplete His-Tag Purification Resin (Roche, SUI).

    Construct:

    Article Title: Ha83, a Chitin Binding Domain Encoding Gene, Is Important to Helicoverpa armigera Nucleopolyhedrovirus Budded Virus Production and Occlusion Body Assembling
    Article Snippet: .. Six truncated Ha83 protein expression vectors (pET-28a-TC151, pET-28a-TC142, pET-28a-TC128, pET-28a-TC118, pET-28a-TC113, and pET-28a-TC71) were constructed by insertion of the truncated ha83 PCR products into Bam HI and Xho I sites of the pET-28a(+) vector (Novagen, GER) with specific primers (Ha83-F used as forward primer, Ha83C151, Ha83C142, Ha83C128, Ha83C118, Ha83C113, and Ha83C71 were used as reverse primers, respectively) ( ). pET-28a-Ha83 which can encode the entire Ha83 was constructed in a similar way with primers Ha83-F and Ha83-R. And pET-28a-ha81, which contains the entire CDs of ha81 gene, was constructed with primers Ha81-F and Ha81-R as described above and used as the negative control plasmid for ha81 sharing a similar base pair number with ha83 . .. The recombinant Ha83, truncated proteins (TC151, TC142, TC128, TC118, TC113, and TC71), as well as the negative control protein Ha81 were then induced by 1 mM isopropyl-β-d-thiogalactoside (IPTG) (Sigma, USA) and purified using affinity chromatography of cOmplete His-Tag Purification Resin (Roche, SUI).

    Produced:

    Article Title: Anti-inflammatory effects of Thymoquinone in activated BV-2 microglia cells
    Article Snippet: .. For iNOS protein expression, cells were fixed in 4% paraformaldehyde/ permeabilized in 0.1% triton X 100 in phosphate buffered saline (PBS) and incubated with anti-iNOS, N-Terminal antibody produced in rabbit (Sigma-Aldrich, St. Louis, MO) for 2 hours at 37°C. .. Samples were washed in PBS and subsequently incubated with anti-rabbit Alexa Fluor® 488 conjugate for two hours at 37°C.

    Incubation:

    Article Title: Anti-inflammatory effects of Thymoquinone in activated BV-2 microglia cells
    Article Snippet: .. For iNOS protein expression, cells were fixed in 4% paraformaldehyde/ permeabilized in 0.1% triton X 100 in phosphate buffered saline (PBS) and incubated with anti-iNOS, N-Terminal antibody produced in rabbit (Sigma-Aldrich, St. Louis, MO) for 2 hours at 37°C. .. Samples were washed in PBS and subsequently incubated with anti-rabbit Alexa Fluor® 488 conjugate for two hours at 37°C.

    Expressing:

    Article Title: Notch1 signaling inhibits growth of EC109 esophageal carcinoma cells through downmodulation of HPV18 E6/E7 gene expression
    Article Snippet: .. Detection of p53 protein expression in EC109 cells by Western blot analysis The proteins from cell lysates of EC109 cells were separated by SDS-polyacrylamide gel electrophoresis and transferred onto PVDF membranes (Millipore, Bedford, MA, USA). .. The blots were incubated with the anti-p53 antibody or the anti-β-actin antibody (1:100 dilution in a blocking solution) at 4 °C overnight, then washed with a solution containing 5% milk, 20 mmol/L Tris-HCl (pH 7.5), 500 mmol/L NaCl, and 0.05% Tween-20 (TBS-T).

    Article Title: Anti-inflammatory effects of Thymoquinone in activated BV-2 microglia cells
    Article Snippet: .. For iNOS protein expression, cells were fixed in 4% paraformaldehyde/ permeabilized in 0.1% triton X 100 in phosphate buffered saline (PBS) and incubated with anti-iNOS, N-Terminal antibody produced in rabbit (Sigma-Aldrich, St. Louis, MO) for 2 hours at 37°C. .. Samples were washed in PBS and subsequently incubated with anti-rabbit Alexa Fluor® 488 conjugate for two hours at 37°C.

    Article Title: Loss of Faap20 causes hematopoietic stem and progenitor cell depletion in mice under genotoxic stress
    Article Snippet: .. FAAP20 protein expression was analyzed in homogenate tissue lysates by western blot using a polyclonal anti-C1orf86 isoform 2 (FAAP20) antibody (Sigma-Aldrich; HPA038829). .. Other antibody used in the experiments: anti-FANCD2 (Novus), anti-phospho-Histone H2A.X (Ser139) (Millipore), and anti-beta-actin (Sigma).

    Article Title: KCa1.1, a calcium-activated potassium channel subunit alpha 1, is targeted by miR-17-5p and modulates cell migration in malignant pleural mesothelioma
    Article Snippet: .. Immunofluorescence staining of KCa1.1 protein expression Cells transfected with KCNMA1-specific siRNA, miR-17-5p mimic or control were fixed in paraformaldehyde solution (4 % v/v in PBS, Sigma, St. Louis, MO, USA) for 15 min, washed three times with PBS and permeabilized with 0.2 % Triton X-100 in PBS for 5 min. ..

    Article Title: Effects of Alginate Concentration and Ovarian Cells on In VitroDevelopment of Mouse Preantral Follicles: A Factorial Study
    Article Snippet: .. Assessment of Cx37 and Cx43 protein expression After deparaffinization and rehydration, antigen retrieval was performed by incubating the sections of the antral follicles in 0.01 M sodium citrate buffer (pH=6.0, Sigma, USA) for 1 hour in a 96°C oven. .. The sections were washed two times in PBS-T, and incubated in 10% goat and donkey serums (Sigma, USA) diluted in PBS for 1 hour at 37°C to block non-specific protein bindings in Cx37 and Cx43 immunostainings, respectively.

    Article Title: N-terminal tyrosine phosphorylation of caveolin-2 negates anti-proliferative effect of transforming growth factor beta in endothelial cells
    Article Snippet: .. We have also tried to determine if N-terminal phosphorylation of Cav-2 affects Col-1 protein expression levels by immunoblotting with the two independent Col-1-specific antibodies (Millipore and Santa Cruz Biotech) but were unable to detect specific signal (data not shown). .. Because of newly described here functional significance of tyrosine phosphorylation in regulating TGF-β signaling and function in ECs, next we have examined if tyrosine 19 and/or 27 phosphorylation of Cav-2 is regulated in ECs.

    Article Title: Blood mononuclear cell gene expression profiles characterize the oxidant, hemolytic, and inflammatory stress of sickle cell disease
    Article Snippet: .. HO-1 protein expression was detected by using 1:1000 dilution of rabbit-anti–HO-1 polyclonal antibody (Calbiochem, La Jolla, CA). .. A 1:5000 dilution of horseradish peroxidase–conjugated goat antirabbit immunoglobulin G (IgG) was used as the secondary antibody (Jackson ImmunoResearch Laboratory, West Grove, PA).

    Article Title: Ha83, a Chitin Binding Domain Encoding Gene, Is Important to Helicoverpa armigera Nucleopolyhedrovirus Budded Virus Production and Occlusion Body Assembling
    Article Snippet: .. Six truncated Ha83 protein expression vectors (pET-28a-TC151, pET-28a-TC142, pET-28a-TC128, pET-28a-TC118, pET-28a-TC113, and pET-28a-TC71) were constructed by insertion of the truncated ha83 PCR products into Bam HI and Xho I sites of the pET-28a(+) vector (Novagen, GER) with specific primers (Ha83-F used as forward primer, Ha83C151, Ha83C142, Ha83C128, Ha83C118, Ha83C113, and Ha83C71 were used as reverse primers, respectively) ( ). pET-28a-Ha83 which can encode the entire Ha83 was constructed in a similar way with primers Ha83-F and Ha83-R. And pET-28a-ha81, which contains the entire CDs of ha81 gene, was constructed with primers Ha81-F and Ha81-R as described above and used as the negative control plasmid for ha81 sharing a similar base pair number with ha83 . .. The recombinant Ha83, truncated proteins (TC151, TC142, TC128, TC118, TC113, and TC71), as well as the negative control protein Ha81 were then induced by 1 mM isopropyl-β-d-thiogalactoside (IPTG) (Sigma, USA) and purified using affinity chromatography of cOmplete His-Tag Purification Resin (Roche, SUI).

    Western Blot:

    Article Title: Notch1 signaling inhibits growth of EC109 esophageal carcinoma cells through downmodulation of HPV18 E6/E7 gene expression
    Article Snippet: .. Detection of p53 protein expression in EC109 cells by Western blot analysis The proteins from cell lysates of EC109 cells were separated by SDS-polyacrylamide gel electrophoresis and transferred onto PVDF membranes (Millipore, Bedford, MA, USA). .. The blots were incubated with the anti-p53 antibody or the anti-β-actin antibody (1:100 dilution in a blocking solution) at 4 °C overnight, then washed with a solution containing 5% milk, 20 mmol/L Tris-HCl (pH 7.5), 500 mmol/L NaCl, and 0.05% Tween-20 (TBS-T).

    Article Title: Loss of Faap20 causes hematopoietic stem and progenitor cell depletion in mice under genotoxic stress
    Article Snippet: .. FAAP20 protein expression was analyzed in homogenate tissue lysates by western blot using a polyclonal anti-C1orf86 isoform 2 (FAAP20) antibody (Sigma-Aldrich; HPA038829). .. Other antibody used in the experiments: anti-FANCD2 (Novus), anti-phospho-Histone H2A.X (Ser139) (Millipore), and anti-beta-actin (Sigma).

    Polymerase Chain Reaction:

    Article Title: Ha83, a Chitin Binding Domain Encoding Gene, Is Important to Helicoverpa armigera Nucleopolyhedrovirus Budded Virus Production and Occlusion Body Assembling
    Article Snippet: .. Six truncated Ha83 protein expression vectors (pET-28a-TC151, pET-28a-TC142, pET-28a-TC128, pET-28a-TC118, pET-28a-TC113, and pET-28a-TC71) were constructed by insertion of the truncated ha83 PCR products into Bam HI and Xho I sites of the pET-28a(+) vector (Novagen, GER) with specific primers (Ha83-F used as forward primer, Ha83C151, Ha83C142, Ha83C128, Ha83C118, Ha83C113, and Ha83C71 were used as reverse primers, respectively) ( ). pET-28a-Ha83 which can encode the entire Ha83 was constructed in a similar way with primers Ha83-F and Ha83-R. And pET-28a-ha81, which contains the entire CDs of ha81 gene, was constructed with primers Ha81-F and Ha81-R as described above and used as the negative control plasmid for ha81 sharing a similar base pair number with ha83 . .. The recombinant Ha83, truncated proteins (TC151, TC142, TC128, TC118, TC113, and TC71), as well as the negative control protein Ha81 were then induced by 1 mM isopropyl-β-d-thiogalactoside (IPTG) (Sigma, USA) and purified using affinity chromatography of cOmplete His-Tag Purification Resin (Roche, SUI).

    Staining:

    Article Title: KCa1.1, a calcium-activated potassium channel subunit alpha 1, is targeted by miR-17-5p and modulates cell migration in malignant pleural mesothelioma
    Article Snippet: .. Immunofluorescence staining of KCa1.1 protein expression Cells transfected with KCNMA1-specific siRNA, miR-17-5p mimic or control were fixed in paraformaldehyde solution (4 % v/v in PBS, Sigma, St. Louis, MO, USA) for 15 min, washed three times with PBS and permeabilized with 0.2 % Triton X-100 in PBS for 5 min. ..

    Immunofluorescence:

    Article Title: KCa1.1, a calcium-activated potassium channel subunit alpha 1, is targeted by miR-17-5p and modulates cell migration in malignant pleural mesothelioma
    Article Snippet: .. Immunofluorescence staining of KCa1.1 protein expression Cells transfected with KCNMA1-specific siRNA, miR-17-5p mimic or control were fixed in paraformaldehyde solution (4 % v/v in PBS, Sigma, St. Louis, MO, USA) for 15 min, washed three times with PBS and permeabilized with 0.2 % Triton X-100 in PBS for 5 min. ..

    Plasmid Preparation:

    Article Title: Ha83, a Chitin Binding Domain Encoding Gene, Is Important to Helicoverpa armigera Nucleopolyhedrovirus Budded Virus Production and Occlusion Body Assembling
    Article Snippet: .. Six truncated Ha83 protein expression vectors (pET-28a-TC151, pET-28a-TC142, pET-28a-TC128, pET-28a-TC118, pET-28a-TC113, and pET-28a-TC71) were constructed by insertion of the truncated ha83 PCR products into Bam HI and Xho I sites of the pET-28a(+) vector (Novagen, GER) with specific primers (Ha83-F used as forward primer, Ha83C151, Ha83C142, Ha83C128, Ha83C118, Ha83C113, and Ha83C71 were used as reverse primers, respectively) ( ). pET-28a-Ha83 which can encode the entire Ha83 was constructed in a similar way with primers Ha83-F and Ha83-R. And pET-28a-ha81, which contains the entire CDs of ha81 gene, was constructed with primers Ha81-F and Ha81-R as described above and used as the negative control plasmid for ha81 sharing a similar base pair number with ha83 . .. The recombinant Ha83, truncated proteins (TC151, TC142, TC128, TC118, TC113, and TC71), as well as the negative control protein Ha81 were then induced by 1 mM isopropyl-β-d-thiogalactoside (IPTG) (Sigma, USA) and purified using affinity chromatography of cOmplete His-Tag Purification Resin (Roche, SUI).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 85
    Millipore bat3 mammalian expression construct
    Modulation of macrophage functions by soluble <t>BAT3.</t> A. J774A.1 cells were first stimulated with IFN-γ (20 ng/ml) for 2 hours and then purified recombinant BAT3 was added into culture at different concentrations. Total nitrite levels in cell culture supernatants were determined after 18 hours using a colorimetric assay kit. **P
    Bat3 Mammalian Expression Construct, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bat3 mammalian expression construct/product/Millipore
    Average 85 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    bat3 mammalian expression construct - by Bioz Stars, 2020-09
    85/100 stars
      Buy from Supplier

    85
    Millipore plasmids expressing cyclin d1
    FBXO31-mediated <t>cyclin</t> D1 degradation occurs through the proteasomal pathway. a , Cyclin D1 levels in SK-MEL-28 cells transduced with a retrovirus expressing FBXO31 or empty vector and treated in the presence or absence of lactacystin. b , Quantitative real-time RT-PCR monitoring cyclin D1 mRNA levels (error bars, s.d., n=3). c , Cyclin D1 levels in SK-MEL-28 cells stably transfected with a plasmid expressing either HA-tagged cyclin D1 or cyclin D1(T286A) and transduced with an FBXO31 retrovirus. d , FACS analysis in cells described in ( d ). e , DNA replication assay (error bars, s.d., n=3).
    Plasmids Expressing Cyclin D1, supplied by Millipore, 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/plasmids expressing cyclin d1/product/Millipore
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    plasmids expressing cyclin d1 - by Bioz Stars, 2020-09
    85/100 stars
      Buy from Supplier

    90
    Millipore e coli protein expression
    Cas9/dCas9 associated toxicity in <t>E.</t> <t>coli</t> strains that are dam + /dcm + or dam − /dcm − .Four E. coli strains were transformed with pRAD1 or pRA-Cas9 or pRA-dCas9 and transformants were elected on carbenicillin selection plates. The CFU were enumerated to determine TE (a). Results are from experiments repeated four times. (b) <t>Protein</t> extracts from E. coli strains, DH5alpha (Lanes A, B,C), JM110 (Lanes D, E, F), JM109 (Lanes G, H, I) and GM2163 (Lanes J, K,L) carrying pRAD1 (Lanes A, D, G and J) or pRA-Cas9 (Lanes B, E, H and K) or pRA-dCas9 (Lanes C, F, I and L) were separated by gel electrophoresis and stained with Coomassie Brilliant Blue (b) or developed for Western blot using Anti-Cas9 antibody (c).
    E Coli Protein Expression, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e coli protein expression/product/Millipore
    Average 90 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    e coli protein expression - by Bioz Stars, 2020-09
    90/100 stars
      Buy from Supplier

    91
    Millipore integrin expression
    A conserved <t>integrin-binding</t> motif exists in the aMPV/B and hMPV F proteins. A , F protein sequences from 12 clinical isolates of aMPV/B and 13 clinical isolates of hMPV were aligned. The RDD motif is highlighted in the boxes. B , the three-dimensional
    Integrin Expression, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/integrin expression/product/Millipore
    Average 91 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    integrin expression - by Bioz Stars, 2020-09
    91/100 stars
      Buy from Supplier

    Image Search Results


    Modulation of macrophage functions by soluble BAT3. A. J774A.1 cells were first stimulated with IFN-γ (20 ng/ml) for 2 hours and then purified recombinant BAT3 was added into culture at different concentrations. Total nitrite levels in cell culture supernatants were determined after 18 hours using a colorimetric assay kit. **P

    Journal: PLoS ONE

    Article Title: BAT3 Regulates Mycobacterium tuberculosis Protein ESAT-6-Mediated Apoptosis of Macrophages

    doi: 10.1371/journal.pone.0040836

    Figure Lengend Snippet: Modulation of macrophage functions by soluble BAT3. A. J774A.1 cells were first stimulated with IFN-γ (20 ng/ml) for 2 hours and then purified recombinant BAT3 was added into culture at different concentrations. Total nitrite levels in cell culture supernatants were determined after 18 hours using a colorimetric assay kit. **P

    Article Snippet: Expression and Purification of Recombinant BAT3 293T cells were transfected with BAT3 mammalian expression construct using Nanojuice transfection kit (EMD Biosciences).

    Techniques: Purification, Recombinant, Cell Culture, Colorimetric Assay

    Regulation of ESAT-6 induced apoptosis by BAT3. A. BMDMs and J774A.1 cells were incubated with different concentrations of ESAT-6 for 24 hours. Cell culture supernatants were collected and subjected to colorimetric caspase-3 assay. Fold change in comparison to unstimulated controls was calculated and plotted in bar graph. *P

    Journal: PLoS ONE

    Article Title: BAT3 Regulates Mycobacterium tuberculosis Protein ESAT-6-Mediated Apoptosis of Macrophages

    doi: 10.1371/journal.pone.0040836

    Figure Lengend Snippet: Regulation of ESAT-6 induced apoptosis by BAT3. A. BMDMs and J774A.1 cells were incubated with different concentrations of ESAT-6 for 24 hours. Cell culture supernatants were collected and subjected to colorimetric caspase-3 assay. Fold change in comparison to unstimulated controls was calculated and plotted in bar graph. *P

    Article Snippet: Expression and Purification of Recombinant BAT3 293T cells were transfected with BAT3 mammalian expression construct using Nanojuice transfection kit (EMD Biosciences).

    Techniques: Incubation, Cell Culture, Caspase-3 Assay

    ESAT-6 induces the expression of BAT3 in macrophages. A. Bone marrow-derived macrophages (BMDM) were incubated with different concentrations of ESAT-6 in Opti-MEM medium for 6 hours. Cell culture supernatants were collected and concentrated. Total protein concentration of different fractions was determined and equal amounts of proteins (25 µg) were loaded on an SDS-PAGE gel. Following electrophoresis and western blotting, the blot was developed using a rabbit polyclonal antibody against BAT3 protein. Histogram plot shows the densitometric quantification of BAT3 protein levels shown in western blot. B. BMDM were incubated with 5 µg/ml of ESAT-6. Nuclear and cytoplasmic extracts were prepared and western blots were developed as mentioned above in figure 3A . Histogram plot shows the densitometric quantification of the change in BAT3 protein levels in nuclear and cytoplasmic extracts. C. Total RNA was isolated from BMDM that were stimulated with ESAT-6 (5 µg/ml) for 6 hours; cDNA was prepared and subjected to real-time PCR for BAT3 gene amplification. ΔΔCT values were normalized to mouse GAPDH gene. D. J774A.1 cells were incubated with 5 µg/ml of ESAT-6 and western blots of nuclear and cytoplasmic fractions were developed as mentioned above in figure 3A . Histogram plot shows the densitometric quantification of the change in BAT3 protein levels in nuclear and cytoplasmic extracts. E. J774A.1 cells were stimulated with ESAT-6 (5 µg/ml) for 6 hours. Total RNA was isolated; cDNA was prepared and subjected to real-time PCR for BAT3 gene amplification. ΔΔCT values were normalized to mouse GAPDH gene. F. J774A.1 cells were incubated with 5 µg/ml of ESAT-6 and culture supernatants were collected at different time points. Heat shock-treated cell supernatants were used as positive control. Western blots were developed as mentioned above in figure 3A . Histogram plot shows the densitometric quantification of BAT3 protein levels shown in western blot. G. J774A.1 cells were incubated with 5 µg/ml of ESAT-6 and cytoplasmic extracts were collected at different time points. The western blots were developed as mentioned above in figure 3A . Histogram plot shows the densitometric quantification of BAT3 protein levels shown in western blot. H. J774A.1 cells were incubated with 5 µg/ml of Ag85B for 9 hours and different cellular fractions (Nuclear extract, Cytoplasmic extract and Supernatants) were collected at different time points. The western blots were developed as mentioned above in figure 3A . The cytoplasmic marker GAPDH and nuclear marker histone H1 served as positive controls for the cytoplasmic extracts and nuclear extracts, respectively, in western blotting experiments.

    Journal: PLoS ONE

    Article Title: BAT3 Regulates Mycobacterium tuberculosis Protein ESAT-6-Mediated Apoptosis of Macrophages

    doi: 10.1371/journal.pone.0040836

    Figure Lengend Snippet: ESAT-6 induces the expression of BAT3 in macrophages. A. Bone marrow-derived macrophages (BMDM) were incubated with different concentrations of ESAT-6 in Opti-MEM medium for 6 hours. Cell culture supernatants were collected and concentrated. Total protein concentration of different fractions was determined and equal amounts of proteins (25 µg) were loaded on an SDS-PAGE gel. Following electrophoresis and western blotting, the blot was developed using a rabbit polyclonal antibody against BAT3 protein. Histogram plot shows the densitometric quantification of BAT3 protein levels shown in western blot. B. BMDM were incubated with 5 µg/ml of ESAT-6. Nuclear and cytoplasmic extracts were prepared and western blots were developed as mentioned above in figure 3A . Histogram plot shows the densitometric quantification of the change in BAT3 protein levels in nuclear and cytoplasmic extracts. C. Total RNA was isolated from BMDM that were stimulated with ESAT-6 (5 µg/ml) for 6 hours; cDNA was prepared and subjected to real-time PCR for BAT3 gene amplification. ΔΔCT values were normalized to mouse GAPDH gene. D. J774A.1 cells were incubated with 5 µg/ml of ESAT-6 and western blots of nuclear and cytoplasmic fractions were developed as mentioned above in figure 3A . Histogram plot shows the densitometric quantification of the change in BAT3 protein levels in nuclear and cytoplasmic extracts. E. J774A.1 cells were stimulated with ESAT-6 (5 µg/ml) for 6 hours. Total RNA was isolated; cDNA was prepared and subjected to real-time PCR for BAT3 gene amplification. ΔΔCT values were normalized to mouse GAPDH gene. F. J774A.1 cells were incubated with 5 µg/ml of ESAT-6 and culture supernatants were collected at different time points. Heat shock-treated cell supernatants were used as positive control. Western blots were developed as mentioned above in figure 3A . Histogram plot shows the densitometric quantification of BAT3 protein levels shown in western blot. G. J774A.1 cells were incubated with 5 µg/ml of ESAT-6 and cytoplasmic extracts were collected at different time points. The western blots were developed as mentioned above in figure 3A . Histogram plot shows the densitometric quantification of BAT3 protein levels shown in western blot. H. J774A.1 cells were incubated with 5 µg/ml of Ag85B for 9 hours and different cellular fractions (Nuclear extract, Cytoplasmic extract and Supernatants) were collected at different time points. The western blots were developed as mentioned above in figure 3A . The cytoplasmic marker GAPDH and nuclear marker histone H1 served as positive controls for the cytoplasmic extracts and nuclear extracts, respectively, in western blotting experiments.

    Article Snippet: Expression and Purification of Recombinant BAT3 293T cells were transfected with BAT3 mammalian expression construct using Nanojuice transfection kit (EMD Biosciences).

    Techniques: Expressing, Derivative Assay, Incubation, Cell Culture, Protein Concentration, SDS Page, Electrophoresis, Western Blot, Isolation, Real-time Polymerase Chain Reaction, Amplification, Positive Control, Marker

    Expression of BAT3 in J774A.1 cells and bone marrow-derived macrophages (BMDM) in response to non-lethal heat shock. A. J774A.1 cells and BMDM were subjected to non-lethal heat shock at 42°C for 10 minutes and rested for 1 hour. Total protein concentration of different fractions was determined and equal amounts of proteins (25 µg) were loaded on SDS-PAGE gels. Following electrophoresis and western blotting, the blots were developed using a rabbit polyclonal antibody against BAT3 protein. The molecular weight of BAT3 was ∼122 kDa. Histogram plot shows the densitometric quantification of BAT3 protein levels shown in western blots. B. Total RNA was isolated from cells; cDNA was prepared and subjected to real-time PCR for BAT3 gene amplification. ΔΔCT values were normalized to mouse GAPDH gene. C. The exosomal pellet (Ex) and soluble fractions (SF), of both J774A.1 cells and bone marrow-derived macrophages (BMDM) culture supernatants were collected. Total 25 µg of protein from each sample was run on a 12% SDS-PAGE gel followed by western blotting to detect BAT3 (upper panel) and HSP70 (lower panel). The cytoplasmic marker GAPDH and nuclear marker histone H1 served as positive controls for the cytoplasmic extracts and nuclear extracts, respectively, in western blotting experiments.

    Journal: PLoS ONE

    Article Title: BAT3 Regulates Mycobacterium tuberculosis Protein ESAT-6-Mediated Apoptosis of Macrophages

    doi: 10.1371/journal.pone.0040836

    Figure Lengend Snippet: Expression of BAT3 in J774A.1 cells and bone marrow-derived macrophages (BMDM) in response to non-lethal heat shock. A. J774A.1 cells and BMDM were subjected to non-lethal heat shock at 42°C for 10 minutes and rested for 1 hour. Total protein concentration of different fractions was determined and equal amounts of proteins (25 µg) were loaded on SDS-PAGE gels. Following electrophoresis and western blotting, the blots were developed using a rabbit polyclonal antibody against BAT3 protein. The molecular weight of BAT3 was ∼122 kDa. Histogram plot shows the densitometric quantification of BAT3 protein levels shown in western blots. B. Total RNA was isolated from cells; cDNA was prepared and subjected to real-time PCR for BAT3 gene amplification. ΔΔCT values were normalized to mouse GAPDH gene. C. The exosomal pellet (Ex) and soluble fractions (SF), of both J774A.1 cells and bone marrow-derived macrophages (BMDM) culture supernatants were collected. Total 25 µg of protein from each sample was run on a 12% SDS-PAGE gel followed by western blotting to detect BAT3 (upper panel) and HSP70 (lower panel). The cytoplasmic marker GAPDH and nuclear marker histone H1 served as positive controls for the cytoplasmic extracts and nuclear extracts, respectively, in western blotting experiments.

    Article Snippet: Expression and Purification of Recombinant BAT3 293T cells were transfected with BAT3 mammalian expression construct using Nanojuice transfection kit (EMD Biosciences).

    Techniques: Expressing, Derivative Assay, Protein Concentration, SDS Page, Electrophoresis, Western Blot, Molecular Weight, Isolation, Real-time Polymerase Chain Reaction, Amplification, Marker

    Inhibition of caspase-3 and proteasome provides protection of BAT3 and BCL-2. J774A.1 cells were pre-incubated with 85 µM zVAD-FMK or 10 µM proteasome inhibitor MG-132 or both for 4 hours and then stimulated with 5 µg/ml of ESAT-6 protein. Cytoplasmic extracts were collected at different time points. Total protein concentration of different fractions was determined and equal amounts of proteins (25 µg) were loaded on an SDS-PAGE gel. Following electrophoresis and western blotting, the blots were developed using a rabbit polyclonal antibody against BAT3 protein ( Figure 6A ) and a mouse monoclonal antibody against BCL-2 protein ( Figure 6B ). All experiments were done with 0.05% DMSO vehicle control and ESAT-6 alone as positive control (data not shown). The cytoplasmic marker GAPDH served as positive control for the western blots of cytoplasmic extracts.

    Journal: PLoS ONE

    Article Title: BAT3 Regulates Mycobacterium tuberculosis Protein ESAT-6-Mediated Apoptosis of Macrophages

    doi: 10.1371/journal.pone.0040836

    Figure Lengend Snippet: Inhibition of caspase-3 and proteasome provides protection of BAT3 and BCL-2. J774A.1 cells were pre-incubated with 85 µM zVAD-FMK or 10 µM proteasome inhibitor MG-132 or both for 4 hours and then stimulated with 5 µg/ml of ESAT-6 protein. Cytoplasmic extracts were collected at different time points. Total protein concentration of different fractions was determined and equal amounts of proteins (25 µg) were loaded on an SDS-PAGE gel. Following electrophoresis and western blotting, the blots were developed using a rabbit polyclonal antibody against BAT3 protein ( Figure 6A ) and a mouse monoclonal antibody against BCL-2 protein ( Figure 6B ). All experiments were done with 0.05% DMSO vehicle control and ESAT-6 alone as positive control (data not shown). The cytoplasmic marker GAPDH served as positive control for the western blots of cytoplasmic extracts.

    Article Snippet: Expression and Purification of Recombinant BAT3 293T cells were transfected with BAT3 mammalian expression construct using Nanojuice transfection kit (EMD Biosciences).

    Techniques: Inhibition, Incubation, Protein Concentration, SDS Page, Electrophoresis, Western Blot, Positive Control, Marker

    Interaction of BAT3 with BCL-2 and expression of BCL-2 in ESAT-6 stimulated J774A.1 cells. A. J774A.1 cells were co-transfected with FLAG-tagged BAT3 plasmid and HIS-tagged BCL-2 plasmid or control vectors and incubated with ESAT-6. Cytoplasmic extracts of the cells were subjected to immunoprecipitation (IP) with antibodies against FLAG peptide or HIS tag, followed by western blotting with respective antibodies as indicated. Total 25 µg of each protein sample was loaded in 12% SDS-PAGE gel for the development of western blot. Upper panel: IP with anti-FLAG antibody and western blot with anti-HIS antibody. The molecular weight of BCL-2 was ∼25 kDa. Lower panel: IP with anti-HIS antibody and western blot with anti-FLAG antibody. The molecular weight of BAT3 was ∼122 kDa. Whole cell lysates of the J774A.1 cells expressing recombinant BAT3 and BCL-2 proteins served as positive control for the western blotting. B. Upper panel: J774A.1 cells were incubated with 5 µg/ml of ESAT-6 and cytoplasmic extracts were collected at different time points. Total protein concentration of different fractions was determined and equal amounts of proteins (25 µg) were loaded on an SDS-PAGE gel. Following electrophoresis and western blotting, the blot was developed using a mouse monoclonal antibody against BCL-2 protein. J774A.1 cells were either transfected with BAT3 plasmid (lower panel) or control vector (middle panel) for 72 hours, incubated with ESAT-6 and cytoplasmic extracts were collected at different time points. Western blots were developed for the BCL-2 protein as described earlier.

    Journal: PLoS ONE

    Article Title: BAT3 Regulates Mycobacterium tuberculosis Protein ESAT-6-Mediated Apoptosis of Macrophages

    doi: 10.1371/journal.pone.0040836

    Figure Lengend Snippet: Interaction of BAT3 with BCL-2 and expression of BCL-2 in ESAT-6 stimulated J774A.1 cells. A. J774A.1 cells were co-transfected with FLAG-tagged BAT3 plasmid and HIS-tagged BCL-2 plasmid or control vectors and incubated with ESAT-6. Cytoplasmic extracts of the cells were subjected to immunoprecipitation (IP) with antibodies against FLAG peptide or HIS tag, followed by western blotting with respective antibodies as indicated. Total 25 µg of each protein sample was loaded in 12% SDS-PAGE gel for the development of western blot. Upper panel: IP with anti-FLAG antibody and western blot with anti-HIS antibody. The molecular weight of BCL-2 was ∼25 kDa. Lower panel: IP with anti-HIS antibody and western blot with anti-FLAG antibody. The molecular weight of BAT3 was ∼122 kDa. Whole cell lysates of the J774A.1 cells expressing recombinant BAT3 and BCL-2 proteins served as positive control for the western blotting. B. Upper panel: J774A.1 cells were incubated with 5 µg/ml of ESAT-6 and cytoplasmic extracts were collected at different time points. Total protein concentration of different fractions was determined and equal amounts of proteins (25 µg) were loaded on an SDS-PAGE gel. Following electrophoresis and western blotting, the blot was developed using a mouse monoclonal antibody against BCL-2 protein. J774A.1 cells were either transfected with BAT3 plasmid (lower panel) or control vector (middle panel) for 72 hours, incubated with ESAT-6 and cytoplasmic extracts were collected at different time points. Western blots were developed for the BCL-2 protein as described earlier.

    Article Snippet: Expression and Purification of Recombinant BAT3 293T cells were transfected with BAT3 mammalian expression construct using Nanojuice transfection kit (EMD Biosciences).

    Techniques: Expressing, Transfection, Plasmid Preparation, Incubation, Immunoprecipitation, Western Blot, SDS Page, Molecular Weight, Recombinant, Positive Control, Protein Concentration, Electrophoresis

    FBXO31-mediated cyclin D1 degradation occurs through the proteasomal pathway. a , Cyclin D1 levels in SK-MEL-28 cells transduced with a retrovirus expressing FBXO31 or empty vector and treated in the presence or absence of lactacystin. b , Quantitative real-time RT-PCR monitoring cyclin D1 mRNA levels (error bars, s.d., n=3). c , Cyclin D1 levels in SK-MEL-28 cells stably transfected with a plasmid expressing either HA-tagged cyclin D1 or cyclin D1(T286A) and transduced with an FBXO31 retrovirus. d , FACS analysis in cells described in ( d ). e , DNA replication assay (error bars, s.d., n=3).

    Journal: Nature

    Article Title: F-Box Protein FBXO31 Mediates Cyclin D1 Degradation to Induce G1 Arrest Following DNA Damage

    doi: 10.1038/nature08011

    Figure Lengend Snippet: FBXO31-mediated cyclin D1 degradation occurs through the proteasomal pathway. a , Cyclin D1 levels in SK-MEL-28 cells transduced with a retrovirus expressing FBXO31 or empty vector and treated in the presence or absence of lactacystin. b , Quantitative real-time RT-PCR monitoring cyclin D1 mRNA levels (error bars, s.d., n=3). c , Cyclin D1 levels in SK-MEL-28 cells stably transfected with a plasmid expressing either HA-tagged cyclin D1 or cyclin D1(T286A) and transduced with an FBXO31 retrovirus. d , FACS analysis in cells described in ( d ). e , DNA replication assay (error bars, s.d., n=3).

    Article Snippet: Cell cycle analysis by two-colour FACS Cells transduced with a retrovirus expressing FBXO31 or empty vector in the presence or absence of plasmids expressing cyclin D1 or cyclin D1(T286A), or cells stably expressing luciferase or cyclin D1 siRNAs, were incubated with BrdU (20 µM) for 4 h, fixed in 70% ethanol and stained sequentially with an α-BrdU mouse antibody (Calbiochem), Alexa 488 conjugated goat α-mouse antibody (Invitrogen) and propidium iodide (Sigma).

    Techniques: Transduction, Expressing, Plasmid Preparation, Quantitative RT-PCR, Stable Transfection, Transfection, FACS

    Ectopic expression of FBXO31 induces G1 arrest and selective degradation of cyclin D1. a , FACS analysis in SK-MEL-28 cells transduced with a retrovirus expressing empty vector or FBXO31 in the absence or presence of nocodazole. b , DNA replication assay, monitored by bromodeoxyuridine (BrdU) incorporation (error bars, s.d., n=3). c–e , Immunoblot analysis showing levels of cyclins ( c ), CDKs ( d ) and CDK inhibitors ( e ) at various time points following transduction with an FBXO31 retrovirus. Tubulin was monitored as a loading control. Ectopic expression of FBXO31 did not induce a DDR ( Supplementary Figure 17 ).

    Journal: Nature

    Article Title: F-Box Protein FBXO31 Mediates Cyclin D1 Degradation to Induce G1 Arrest Following DNA Damage

    doi: 10.1038/nature08011

    Figure Lengend Snippet: Ectopic expression of FBXO31 induces G1 arrest and selective degradation of cyclin D1. a , FACS analysis in SK-MEL-28 cells transduced with a retrovirus expressing empty vector or FBXO31 in the absence or presence of nocodazole. b , DNA replication assay, monitored by bromodeoxyuridine (BrdU) incorporation (error bars, s.d., n=3). c–e , Immunoblot analysis showing levels of cyclins ( c ), CDKs ( d ) and CDK inhibitors ( e ) at various time points following transduction with an FBXO31 retrovirus. Tubulin was monitored as a loading control. Ectopic expression of FBXO31 did not induce a DDR ( Supplementary Figure 17 ).

    Article Snippet: Cell cycle analysis by two-colour FACS Cells transduced with a retrovirus expressing FBXO31 or empty vector in the presence or absence of plasmids expressing cyclin D1 or cyclin D1(T286A), or cells stably expressing luciferase or cyclin D1 siRNAs, were incubated with BrdU (20 µM) for 4 h, fixed in 70% ethanol and stained sequentially with an α-BrdU mouse antibody (Calbiochem), Alexa 488 conjugated goat α-mouse antibody (Invitrogen) and propidium iodide (Sigma).

    Techniques: Expressing, FACS, Transduction, Plasmid Preparation, BrdU Incorporation Assay

    FBXO31 interacts with and directs ubiquitination of cyclin D1. a , Co-immunoprecipitation of FBXO31 with cyclin D1 and SCF complex components. The FBXO31-cyclin D1 interaction was specific ( Supplementary Fig. 18 ) and predominantly cytoplasmic ( Supplementary Fig. 19 ). b , Co-immunoprecipitation of endogenous FBXO31 and cyclin D1. c , Cyclin D1 levels in SK-MEL-28 cells ectopically expressing vector, FBXO31 or FBXO31ΔF. d , In vivo ubiquitination assay. Polyubiquitinated cyclin D1 was detected by immunoprecipitation of Flag-tagged ubiquitin followed by immunoblotting for HA-cyclin D1. e , In vitro ubiquitination assay. Immunopurified SCF complexes were incubated with GST-cyclin D1, Erk2, E1, E2, ATP and ubiquitin.

    Journal: Nature

    Article Title: F-Box Protein FBXO31 Mediates Cyclin D1 Degradation to Induce G1 Arrest Following DNA Damage

    doi: 10.1038/nature08011

    Figure Lengend Snippet: FBXO31 interacts with and directs ubiquitination of cyclin D1. a , Co-immunoprecipitation of FBXO31 with cyclin D1 and SCF complex components. The FBXO31-cyclin D1 interaction was specific ( Supplementary Fig. 18 ) and predominantly cytoplasmic ( Supplementary Fig. 19 ). b , Co-immunoprecipitation of endogenous FBXO31 and cyclin D1. c , Cyclin D1 levels in SK-MEL-28 cells ectopically expressing vector, FBXO31 or FBXO31ΔF. d , In vivo ubiquitination assay. Polyubiquitinated cyclin D1 was detected by immunoprecipitation of Flag-tagged ubiquitin followed by immunoblotting for HA-cyclin D1. e , In vitro ubiquitination assay. Immunopurified SCF complexes were incubated with GST-cyclin D1, Erk2, E1, E2, ATP and ubiquitin.

    Article Snippet: Cell cycle analysis by two-colour FACS Cells transduced with a retrovirus expressing FBXO31 or empty vector in the presence or absence of plasmids expressing cyclin D1 or cyclin D1(T286A), or cells stably expressing luciferase or cyclin D1 siRNAs, were incubated with BrdU (20 µM) for 4 h, fixed in 70% ethanol and stained sequentially with an α-BrdU mouse antibody (Calbiochem), Alexa 488 conjugated goat α-mouse antibody (Invitrogen) and propidium iodide (Sigma).

    Techniques: Immunoprecipitation, Expressing, Plasmid Preparation, In Vivo, Ubiquitin Assay, In Vitro, Incubation

    Cell cycle arrest following DNA damage requires ATM-mediated induction of FBXO31. a , Immunoblot analysis of FBXO31 following γ-irradiation. For comparison to ectopically expressed FBXO31 levels see Supplementary Fig. 20 . b , Cyclin D1 levels in cells expressing a NS or FBXO31 shRNA following γ-irradiation. c , FACS analysis. d , (Top) Putative ATM sites in FBXO31. (Bottom) In vitro phosphorylation of GST-FBXO31 fusion proteins harbouring wild type (WT) or mutated (SDM) SQ sites. ATM-WT, wild type ATM; ATM-KD, kinase dead ATM mutant. e–f , Levels of ectopically expressed WT or mutant FBXO31 following γ-irradiation (e) or ectopic ATM expression (f). g , FBXO31, cyclin D1, ATM and phosphorylated ATM in γ-irradiated cells expressing a NS or ATM shRNA. h , Colony formation assay in untreated or γ-irradiated cells. i , FBXO31 and cyclin D1 levels following treatment with DNA damaging agents. j , Model.

    Journal: Nature

    Article Title: F-Box Protein FBXO31 Mediates Cyclin D1 Degradation to Induce G1 Arrest Following DNA Damage

    doi: 10.1038/nature08011

    Figure Lengend Snippet: Cell cycle arrest following DNA damage requires ATM-mediated induction of FBXO31. a , Immunoblot analysis of FBXO31 following γ-irradiation. For comparison to ectopically expressed FBXO31 levels see Supplementary Fig. 20 . b , Cyclin D1 levels in cells expressing a NS or FBXO31 shRNA following γ-irradiation. c , FACS analysis. d , (Top) Putative ATM sites in FBXO31. (Bottom) In vitro phosphorylation of GST-FBXO31 fusion proteins harbouring wild type (WT) or mutated (SDM) SQ sites. ATM-WT, wild type ATM; ATM-KD, kinase dead ATM mutant. e–f , Levels of ectopically expressed WT or mutant FBXO31 following γ-irradiation (e) or ectopic ATM expression (f). g , FBXO31, cyclin D1, ATM and phosphorylated ATM in γ-irradiated cells expressing a NS or ATM shRNA. h , Colony formation assay in untreated or γ-irradiated cells. i , FBXO31 and cyclin D1 levels following treatment with DNA damaging agents. j , Model.

    Article Snippet: Cell cycle analysis by two-colour FACS Cells transduced with a retrovirus expressing FBXO31 or empty vector in the presence or absence of plasmids expressing cyclin D1 or cyclin D1(T286A), or cells stably expressing luciferase or cyclin D1 siRNAs, were incubated with BrdU (20 µM) for 4 h, fixed in 70% ethanol and stained sequentially with an α-BrdU mouse antibody (Calbiochem), Alexa 488 conjugated goat α-mouse antibody (Invitrogen) and propidium iodide (Sigma).

    Techniques: Irradiation, Expressing, shRNA, FACS, In Vitro, Mutagenesis, Colony Assay

    Cas9/dCas9 associated toxicity in E. coli strains that are dam + /dcm + or dam − /dcm − .Four E. coli strains were transformed with pRAD1 or pRA-Cas9 or pRA-dCas9 and transformants were elected on carbenicillin selection plates. The CFU were enumerated to determine TE (a). Results are from experiments repeated four times. (b) Protein extracts from E. coli strains, DH5alpha (Lanes A, B,C), JM110 (Lanes D, E, F), JM109 (Lanes G, H, I) and GM2163 (Lanes J, K,L) carrying pRAD1 (Lanes A, D, G and J) or pRA-Cas9 (Lanes B, E, H and K) or pRA-dCas9 (Lanes C, F, I and L) were separated by gel electrophoresis and stained with Coomassie Brilliant Blue (b) or developed for Western blot using Anti-Cas9 antibody (c).

    Journal: bioRxiv

    Article Title: Determination of Cas9/dCas9 associated toxicity in microbes

    doi: 10.1101/848135

    Figure Lengend Snippet: Cas9/dCas9 associated toxicity in E. coli strains that are dam + /dcm + or dam − /dcm − .Four E. coli strains were transformed with pRAD1 or pRA-Cas9 or pRA-dCas9 and transformants were elected on carbenicillin selection plates. The CFU were enumerated to determine TE (a). Results are from experiments repeated four times. (b) Protein extracts from E. coli strains, DH5alpha (Lanes A, B,C), JM110 (Lanes D, E, F), JM109 (Lanes G, H, I) and GM2163 (Lanes J, K,L) carrying pRAD1 (Lanes A, D, G and J) or pRA-Cas9 (Lanes B, E, H and K) or pRA-dCas9 (Lanes C, F, I and L) were separated by gel electrophoresis and stained with Coomassie Brilliant Blue (b) or developed for Western blot using Anti-Cas9 antibody (c).

    Article Snippet: Expression of Cas9 and dCas9 in E. coli Expression of Cas9 and dCas9 proteins was determined by separation of protein extracts of E. coli strains bearing the pRAD1, or pRA-cas9 and pRA-dcas9 by electrophoresis on a 10% denaturing polyacrylamide gel.

    Techniques: Transformation Assay, Selection, Nucleic Acid Electrophoresis, Staining, Western Blot

    A conserved integrin-binding motif exists in the aMPV/B and hMPV F proteins. A , F protein sequences from 12 clinical isolates of aMPV/B and 13 clinical isolates of hMPV were aligned. The RDD motif is highlighted in the boxes. B , the three-dimensional

    Journal: The Journal of Biological Chemistry

    Article Title: Integrin αvβ1 Modulation Affects Subtype B Avian Metapneumovirus Fusion Protein-mediated Cell-Cell Fusion and Virus Infection *

    doi: 10.1074/jbc.M115.711382

    Figure Lengend Snippet: A conserved integrin-binding motif exists in the aMPV/B and hMPV F proteins. A , F protein sequences from 12 clinical isolates of aMPV/B and 13 clinical isolates of hMPV were aligned. The RDD motif is highlighted in the boxes. B , the three-dimensional

    Article Snippet: As a reference control for the analysis of F protein and integrin expression on the cell membrane, Na+ /K+ -ATPase expression on the cell membrane was analyzed using an anti-Na+ /K+ -ATPase β-1 antibody (monoclonal antibody, catalogue number: 05-382, lot number: 2685213, Millipore).

    Techniques: Binding Assay

    Knockdown of integrin αv- and β1-specific siRNAs inhibits the fusogenic activity of the aMPV/B F protein and the replication of aMPV/B. A, siRNAs targeting monkey integrin αv or β1, mouse integrin αv or β1,

    Journal: The Journal of Biological Chemistry

    Article Title: Integrin αvβ1 Modulation Affects Subtype B Avian Metapneumovirus Fusion Protein-mediated Cell-Cell Fusion and Virus Infection *

    doi: 10.1074/jbc.M115.711382

    Figure Lengend Snippet: Knockdown of integrin αv- and β1-specific siRNAs inhibits the fusogenic activity of the aMPV/B F protein and the replication of aMPV/B. A, siRNAs targeting monkey integrin αv or β1, mouse integrin αv or β1,

    Article Snippet: As a reference control for the analysis of F protein and integrin expression on the cell membrane, Na+ /K+ -ATPase expression on the cell membrane was analyzed using an anti-Na+ /K+ -ATPase β-1 antibody (monoclonal antibody, catalogue number: 05-382, lot number: 2685213, Millipore).

    Techniques: Activity Assay

    Overexpression of integrin αv and β1 enhances the fusogenic activity of the aMPV/B F protein and the replication of aMPV/B. A–D , cells were transfected with plasmid DNAs expressing integrin αv and β1 from different

    Journal: The Journal of Biological Chemistry

    Article Title: Integrin αvβ1 Modulation Affects Subtype B Avian Metapneumovirus Fusion Protein-mediated Cell-Cell Fusion and Virus Infection *

    doi: 10.1074/jbc.M115.711382

    Figure Lengend Snippet: Overexpression of integrin αv and β1 enhances the fusogenic activity of the aMPV/B F protein and the replication of aMPV/B. A–D , cells were transfected with plasmid DNAs expressing integrin αv and β1 from different

    Article Snippet: As a reference control for the analysis of F protein and integrin expression on the cell membrane, Na+ /K+ -ATPase expression on the cell membrane was analyzed using an anti-Na+ /K+ -ATPase β-1 antibody (monoclonal antibody, catalogue number: 05-382, lot number: 2685213, Millipore).

    Techniques: Over Expression, Activity Assay, Transfection, Plasmid Preparation, Expressing

    Overexpression of integrin αv and β1 enhances the binding activity of the aMPV/B F protein and infection of aMPV/B. A , the binding activity of the aMPV/B F protein and infection with aMPV/B in CHO cells overexpressing the integrins αv

    Journal: The Journal of Biological Chemistry

    Article Title: Integrin αvβ1 Modulation Affects Subtype B Avian Metapneumovirus Fusion Protein-mediated Cell-Cell Fusion and Virus Infection *

    doi: 10.1074/jbc.M115.711382

    Figure Lengend Snippet: Overexpression of integrin αv and β1 enhances the binding activity of the aMPV/B F protein and infection of aMPV/B. A , the binding activity of the aMPV/B F protein and infection with aMPV/B in CHO cells overexpressing the integrins αv

    Article Snippet: As a reference control for the analysis of F protein and integrin expression on the cell membrane, Na+ /K+ -ATPase expression on the cell membrane was analyzed using an anti-Na+ /K+ -ATPase β-1 antibody (monoclonal antibody, catalogue number: 05-382, lot number: 2685213, Millipore).

    Techniques: Over Expression, Binding Assay, Activity Assay, Infection

    Antibodies specific for αv and β1 integrin block the fusogenic activity of the aMPV/B F protein and aMPV/B replication. A and B , in the presence of integrin-specific antibodies, aMPV/B F protein fusogenic activities were measured by syncytium

    Journal: The Journal of Biological Chemistry

    Article Title: Integrin αvβ1 Modulation Affects Subtype B Avian Metapneumovirus Fusion Protein-mediated Cell-Cell Fusion and Virus Infection *

    doi: 10.1074/jbc.M115.711382

    Figure Lengend Snippet: Antibodies specific for αv and β1 integrin block the fusogenic activity of the aMPV/B F protein and aMPV/B replication. A and B , in the presence of integrin-specific antibodies, aMPV/B F protein fusogenic activities were measured by syncytium

    Article Snippet: As a reference control for the analysis of F protein and integrin expression on the cell membrane, Na+ /K+ -ATPase expression on the cell membrane was analyzed using an anti-Na+ /K+ -ATPase β-1 antibody (monoclonal antibody, catalogue number: 05-382, lot number: 2685213, Millipore).

    Techniques: Blocking Assay, Activity Assay