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  • 99
    ATCC ma104 cells
    Electron microscopy of RV-infected cells treated with ML. High-definition electron microscopy of noninfected (NI) and RV-infected (OSU; MOI, 100 VFU/ml) <t>MA104</t> cells untreated (DMSO) or treated with ML (20 μM) from 1 hpi. At 6 hpi, the cells were fixed with glutaraldehyde and processed for transmission electron microscopy. V, viroplasms; Nu, nucleus, ER, endoplasmic reticulum; Gg, Golgi complex; Vc, vacuoles; P h , phagosomes; CM, cell membrane; the thin arrows indicate the endoplasmic reticulum membrane surrounding viroplasms; the large arrowheads indicate viral particles.
    Ma104 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 227 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher micro bca assay
    M s self-assembles. M i and M s were incubated at 500 nM or with equivalent amounts (monomer equivalent) of dimer and trimer for various times prior to resolution by <t>SEC.</t> Assemblies were monitored by reading the absorbance of fractions using micro <t>BCA</t> assay. ( A ) M i showed no self-association. ( B ) M s exhibited self-association over time. ( C,D ) Dimer and trimer were stable over time. ( E,F ) M i does not react with dimer or trimer to form larger assemblies. ( G,H ) M s reacts with dimer and trimer to form larger assemblies. See Figure 5—source data 1 . 10.7554/eLife.36584.012 Data for M s self-assembly.
    Micro Bca Assay, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3841 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore 3 methyladenine 3 ma
    Pharmacological and gene silencing of autophagy markers blocks cocaine-mediated mitophagy. (a–f) Representative western blots showing expression of mitophagy markers such as PINK1 (a), PRKN (b), and DNM1L (c) and autophagy markers such as BECN1 (d), MAP1LC3B-II (e), and SQSTM1 (f) in mPMs pretreated with either 5 mM of <t>3-methyladenine</t> (3-MA) or 100 nM of wortmannin for 1 h following exposure of cells to 10 μM cocaine for 24 h. (g–l) Representative western blots showing the expression of PINK1 (g), PRKN (h), and DNM1L (i), BECN1 (j), MAP1LC3B-II (k), and SQSTM1 (l) in mPMs transfected with BECN1 siRNA and scrambled siRNA following exposure of cells to 10 μM cocaine for 24 h. ACTB was probed as a protein loading control for all experiments. The data are presented as mean±SEM from six independent experiments. Non-parametric Kruskal-Wallis One-way ANOVA followed by Dunn’s post hoc test was used to determine the statistical significance among multiple groups. *, P
    3 Methyladenine 3 Ma, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 290 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Selleck Chemicals 3 methyladenine
    Effect of curcumin treatment on the cell cycle and apoptosis of NSCs in suspended state. NSCs were treated with control, curcumin and 3MA for 72 h. β-actin was used as the loading control. The relative level of protein was quantified by ImageJ. Experiments were repeated three times. NSC, neural stem cells; 3MA, <t>3-methyladenine.</t>
    3 Methyladenine, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 99/100, based on 185 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Janssen dragon durey ma
    Effect of curcumin treatment on the cell cycle and apoptosis of NSCs in suspended state. NSCs were treated with control, curcumin and 3MA for 72 h. β-actin was used as the loading control. The relative level of protein was quantified by ImageJ. Experiments were repeated three times. NSC, neural stem cells; 3MA, <t>3-methyladenine.</t>
    Dragon Durey Ma, supplied by Janssen, used in various techniques. Bioz Stars score: 90/100, based on 97 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Vollrath landolt ma
    Effect of curcumin treatment on the cell cycle and apoptosis of NSCs in suspended state. NSCs were treated with control, curcumin and 3MA for 72 h. β-actin was used as the loading control. The relative level of protein was quantified by ImageJ. Experiments were repeated three times. NSC, neural stem cells; 3MA, <t>3-methyladenine.</t>
    Landolt Ma, supplied by Vollrath, used in various techniques. Bioz Stars score: 91/100, based on 83 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Staples ma 11 2 11 ma
    Preparation and characterization of 2D net <t>-poly(MA-11-2-11-MA).</t> ( A ) Schematic illustration of the molecular structure of gemini monomer MA-11-2-11-MA and 2D net- poly(MA-11-2-11-MA). ( B ) AFM image and height analysis of the 2D net- poly(MA-11-2-11-MA) on the silicon substrate. Rq, root mean square roughness. ( C ) Simulation of the bilayer thickness with vertically arranged MA-11-2-11-MA monomers using Materials Studio. ( D ) TEM image of the freestanding film of 2D net -poly(MA-11-2-11-MA).
    Ma 11 2 11 Ma, supplied by Staples, used in various techniques. Bioz Stars score: 94/100, based on 58 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    ChromoTek gfp trap ma beads
    TCP8, TCP14, and TCP15 interact with <t>NPR1.</t> (A) Interactions between NPR1 with TCP8, TCP14, and TCP15 in yeast two-hybrid assays. Diploid yeast cells were serially diluted at OD 600 = 1.0, 0.1, and 0.01. Aliquots of 10 μL of each dilution were plated on double synthetic dropout control plates without leucine and tryptophan (-LW) and selective triple synthetic dropout plates without leucine, tryptophan, and histidine with 1 mM 3-amino-1,2,4-triazole (–LWH + 1 mM 3AT). Photographs were taken 5 days after plating. EV, Empty vector; BD, GAL4 DNA-binding domain; AD, GAL4 activation domain. (B) Co-immunoprecipitation (Co-IP) of NPR1 with TCP8 in Nicotiana benthamiana . <t>NPR1-GFP</t> and FLAG-TCP8 under control of the cauliflower mosaic virus (CMV) 35S promoter were transiently co-expressed in N . benthamiana by agroinfiltration. Total protein extracts were immunoprecipitated with anti-FLAG M2 magnetic beads. The input and immunoprecipitated protein were analyzed by immunoblot using the anti-FLAG and anti-GFP antibodies. kDa, kilodaltons. (C,D) Co-IP of TCP14 and TCP15 with NPR1 in N . benthamiana . Constitutively expressed NPR1-GFP was transiently co-expressed with HA-TCP14 (C) and HA-TCP15 (D) under control of the CMV 35S promoter in N . benthamiana by agroinfiltration. Total protein extracts were immunoprecipitated using anti-GFP trap beads. The input and immuno-precipitated proteins were analyzed by immunoblot using anti-HA and anti-GFP antibodies.
    Gfp Trap Ma Beads, supplied by ChromoTek, used in various techniques. Bioz Stars score: 89/100, based on 253 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Bio-Rad ma cm2
    TCP8, TCP14, and TCP15 interact with <t>NPR1.</t> (A) Interactions between NPR1 with TCP8, TCP14, and TCP15 in yeast two-hybrid assays. Diploid yeast cells were serially diluted at OD 600 = 1.0, 0.1, and 0.01. Aliquots of 10 μL of each dilution were plated on double synthetic dropout control plates without leucine and tryptophan (-LW) and selective triple synthetic dropout plates without leucine, tryptophan, and histidine with 1 mM 3-amino-1,2,4-triazole (–LWH + 1 mM 3AT). Photographs were taken 5 days after plating. EV, Empty vector; BD, GAL4 DNA-binding domain; AD, GAL4 activation domain. (B) Co-immunoprecipitation (Co-IP) of NPR1 with TCP8 in Nicotiana benthamiana . <t>NPR1-GFP</t> and FLAG-TCP8 under control of the cauliflower mosaic virus (CMV) 35S promoter were transiently co-expressed in N . benthamiana by agroinfiltration. Total protein extracts were immunoprecipitated with anti-FLAG M2 magnetic beads. The input and immunoprecipitated protein were analyzed by immunoblot using the anti-FLAG and anti-GFP antibodies. kDa, kilodaltons. (C,D) Co-IP of TCP14 and TCP15 with NPR1 in N . benthamiana . Constitutively expressed NPR1-GFP was transiently co-expressed with HA-TCP14 (C) and HA-TCP15 (D) under control of the CMV 35S promoter in N . benthamiana by agroinfiltration. Total protein extracts were immunoprecipitated using anti-GFP trap beads. The input and immuno-precipitated proteins were analyzed by immunoblot using anti-HA and anti-GFP antibodies.
    Ma Cm2, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 279 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    DuPont de Nemours ma jz
    TCP8, TCP14, and TCP15 interact with <t>NPR1.</t> (A) Interactions between NPR1 with TCP8, TCP14, and TCP15 in yeast two-hybrid assays. Diploid yeast cells were serially diluted at OD 600 = 1.0, 0.1, and 0.01. Aliquots of 10 μL of each dilution were plated on double synthetic dropout control plates without leucine and tryptophan (-LW) and selective triple synthetic dropout plates without leucine, tryptophan, and histidine with 1 mM 3-amino-1,2,4-triazole (–LWH + 1 mM 3AT). Photographs were taken 5 days after plating. EV, Empty vector; BD, GAL4 DNA-binding domain; AD, GAL4 activation domain. (B) Co-immunoprecipitation (Co-IP) of NPR1 with TCP8 in Nicotiana benthamiana . <t>NPR1-GFP</t> and FLAG-TCP8 under control of the cauliflower mosaic virus (CMV) 35S promoter were transiently co-expressed in N . benthamiana by agroinfiltration. Total protein extracts were immunoprecipitated with anti-FLAG M2 magnetic beads. The input and immunoprecipitated protein were analyzed by immunoblot using the anti-FLAG and anti-GFP antibodies. kDa, kilodaltons. (C,D) Co-IP of TCP14 and TCP15 with NPR1 in N . benthamiana . Constitutively expressed NPR1-GFP was transiently co-expressed with HA-TCP14 (C) and HA-TCP15 (D) under control of the CMV 35S promoter in N . benthamiana by agroinfiltration. Total protein extracts were immunoprecipitated using anti-GFP trap beads. The input and immuno-precipitated proteins were analyzed by immunoblot using anti-HA and anti-GFP antibodies.
    Ma Jz, supplied by DuPont de Nemours, used in various techniques. Bioz Stars score: 90/100, based on 123 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Abcam npm1 ma
    INPP4B is upregulated by <t>NPM1-mA</t> in leukemia cells via ERK/Ets-1 signaling. ( a ) qRT-PCR analysis of INPP4B mRNA expression, ( b ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. ( c ) qRT-PCR analysis of INPP4B mRNA expression, ( d ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the THP-1 and K562 cells transduced with the plasmids expressing NPM1-wt or NPM1-mA. ( e ) qRT-PCR analysis of INPP4B mRNA expression, ( f ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3, p-Ets-1 and Ets-1 from the OCI-AML3 cells transfected with the control siRNA or siEts-1. ( g ) Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 and SGK3, ( h ) qRT-PCR analysis of INPP4B mRNA expression from the OCI-AML3 cells treated with different concentration of PD98059 (0, 10, 20 and 40 μM). i Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. Proteins were quantified using image software and normalized against β-actin. Data were represented as mean ± s.d. of three individual experiments. * p
    Npm1 Ma, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 47 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Greenlee barhoover ma
    INPP4B is upregulated by <t>NPM1-mA</t> in leukemia cells via ERK/Ets-1 signaling. ( a ) qRT-PCR analysis of INPP4B mRNA expression, ( b ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. ( c ) qRT-PCR analysis of INPP4B mRNA expression, ( d ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the THP-1 and K562 cells transduced with the plasmids expressing NPM1-wt or NPM1-mA. ( e ) qRT-PCR analysis of INPP4B mRNA expression, ( f ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3, p-Ets-1 and Ets-1 from the OCI-AML3 cells transfected with the control siRNA or siEts-1. ( g ) Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 and SGK3, ( h ) qRT-PCR analysis of INPP4B mRNA expression from the OCI-AML3 cells treated with different concentration of PD98059 (0, 10, 20 and 40 μM). i Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. Proteins were quantified using image software and normalized against β-actin. Data were represented as mean ± s.d. of three individual experiments. * p
    Barhoover Ma, supplied by Greenlee, used in various techniques. Bioz Stars score: 88/100, based on 84 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    ATCC a castellanii atcc 50370
    INPP4B is upregulated by <t>NPM1-mA</t> in leukemia cells via ERK/Ets-1 signaling. ( a ) qRT-PCR analysis of INPP4B mRNA expression, ( b ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. ( c ) qRT-PCR analysis of INPP4B mRNA expression, ( d ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the THP-1 and K562 cells transduced with the plasmids expressing NPM1-wt or NPM1-mA. ( e ) qRT-PCR analysis of INPP4B mRNA expression, ( f ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3, p-Ets-1 and Ets-1 from the OCI-AML3 cells transfected with the control siRNA or siEts-1. ( g ) Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 and SGK3, ( h ) qRT-PCR analysis of INPP4B mRNA expression from the OCI-AML3 cells treated with different concentration of PD98059 (0, 10, 20 and 40 μM). i Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. Proteins were quantified using image software and normalized against β-actin. Data were represented as mean ± s.d. of three individual experiments. * p
    A Castellanii Atcc 50370, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 60 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    RStudio ma url
    INPP4B is upregulated by <t>NPM1-mA</t> in leukemia cells via ERK/Ets-1 signaling. ( a ) qRT-PCR analysis of INPP4B mRNA expression, ( b ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. ( c ) qRT-PCR analysis of INPP4B mRNA expression, ( d ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the THP-1 and K562 cells transduced with the plasmids expressing NPM1-wt or NPM1-mA. ( e ) qRT-PCR analysis of INPP4B mRNA expression, ( f ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3, p-Ets-1 and Ets-1 from the OCI-AML3 cells transfected with the control siRNA or siEts-1. ( g ) Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 and SGK3, ( h ) qRT-PCR analysis of INPP4B mRNA expression from the OCI-AML3 cells treated with different concentration of PD98059 (0, 10, 20 and 40 μM). i Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. Proteins were quantified using image software and normalized against β-actin. Data were represented as mean ± s.d. of three individual experiments. * p
    Ma Url, supplied by RStudio, used in various techniques. Bioz Stars score: 91/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Greiner Bio contreras ma
    INPP4B is upregulated by <t>NPM1-mA</t> in leukemia cells via ERK/Ets-1 signaling. ( a ) qRT-PCR analysis of INPP4B mRNA expression, ( b ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. ( c ) qRT-PCR analysis of INPP4B mRNA expression, ( d ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the THP-1 and K562 cells transduced with the plasmids expressing NPM1-wt or NPM1-mA. ( e ) qRT-PCR analysis of INPP4B mRNA expression, ( f ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3, p-Ets-1 and Ets-1 from the OCI-AML3 cells transfected with the control siRNA or siEts-1. ( g ) Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 and SGK3, ( h ) qRT-PCR analysis of INPP4B mRNA expression from the OCI-AML3 cells treated with different concentration of PD98059 (0, 10, 20 and 40 μM). i Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. Proteins were quantified using image software and normalized against β-actin. Data were represented as mean ± s.d. of three individual experiments. * p
    Contreras Ma, supplied by Greiner Bio, used in various techniques. Bioz Stars score: 88/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    ATCC s avermitilis atcc 31267
    Effect of SAV7472-SAV7473 overexpression on avermectin production in S. <t>avermitilis</t> ATCC 31267. The strains (left to right) are wild-type ATCC 31267, plasmid control strain ATCC 31267(pKC1139), and SAV7472-SAV7473 overexpressing transformants 31267(pKCE7273)-1,
    S Avermitilis Atcc 31267, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 87 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Hallwachs smith ma
    Effect of SAV7472-SAV7473 overexpression on avermectin production in S. <t>avermitilis</t> ATCC 31267. The strains (left to right) are wild-type ATCC 31267, plasmid control strain ATCC 31267(pKC1139), and SAV7472-SAV7473 overexpressing transformants 31267(pKCE7273)-1,
    Smith Ma, supplied by Hallwachs, used in various techniques. Bioz Stars score: 95/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher carbopac ma1 column
    Effect of SAV7472-SAV7473 overexpression on avermectin production in S. <t>avermitilis</t> ATCC 31267. The strains (left to right) are wild-type ATCC 31267, plasmid control strain ATCC 31267(pKC1139), and SAV7472-SAV7473 overexpressing transformants 31267(pKCE7273)-1,
    Carbopac Ma1 Column, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 65 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Electron microscopy of RV-infected cells treated with ML. High-definition electron microscopy of noninfected (NI) and RV-infected (OSU; MOI, 100 VFU/ml) MA104 cells untreated (DMSO) or treated with ML (20 μM) from 1 hpi. At 6 hpi, the cells were fixed with glutaraldehyde and processed for transmission electron microscopy. V, viroplasms; Nu, nucleus, ER, endoplasmic reticulum; Gg, Golgi complex; Vc, vacuoles; P h , phagosomes; CM, cell membrane; the thin arrows indicate the endoplasmic reticulum membrane surrounding viroplasms; the large arrowheads indicate viral particles.

    Journal: Journal of Virology

    Article Title: Identification of a Small Molecule That Compromises the Structural Integrity of Viroplasms and Rotavirus Double-Layered Particles

    doi: 10.1128/JVI.01943-17

    Figure Lengend Snippet: Electron microscopy of RV-infected cells treated with ML. High-definition electron microscopy of noninfected (NI) and RV-infected (OSU; MOI, 100 VFU/ml) MA104 cells untreated (DMSO) or treated with ML (20 μM) from 1 hpi. At 6 hpi, the cells were fixed with glutaraldehyde and processed for transmission electron microscopy. V, viroplasms; Nu, nucleus, ER, endoplasmic reticulum; Gg, Golgi complex; Vc, vacuoles; P h , phagosomes; CM, cell membrane; the thin arrows indicate the endoplasmic reticulum membrane surrounding viroplasms; the large arrowheads indicate viral particles.

    Article Snippet: MA104 cells (embryonic African green monkey kidney cells; ATCC CRL-2378) were grown in Dulbecco's modified Eagle's medium (DMEM) (Life Technologies) containing 10% fetal bovine serum (Life Technologies) and 50 μg/ml gentamicin (Biochrom AG).

    Techniques: Electron Microscopy, Infection, Transmission Assay

    NSP5 dephosphorylation caused by ML-mediated viroplasm disruption. Shown are Western blot and confocal immunofluorescence analyses with the indicated antibodies of OSU-infected (25 VFU/cell) MA104 cells treated with 10 μM ML and/or 0.5 μM okadaic acid (OA) or DMSO for the indicated times. Scale bars, 5 μm.

    Journal: Journal of Virology

    Article Title: Identification of a Small Molecule That Compromises the Structural Integrity of Viroplasms and Rotavirus Double-Layered Particles

    doi: 10.1128/JVI.01943-17

    Figure Lengend Snippet: NSP5 dephosphorylation caused by ML-mediated viroplasm disruption. Shown are Western blot and confocal immunofluorescence analyses with the indicated antibodies of OSU-infected (25 VFU/cell) MA104 cells treated with 10 μM ML and/or 0.5 μM okadaic acid (OA) or DMSO for the indicated times. Scale bars, 5 μm.

    Article Snippet: MA104 cells (embryonic African green monkey kidney cells; ATCC CRL-2378) were grown in Dulbecco's modified Eagle's medium (DMEM) (Life Technologies) containing 10% fetal bovine serum (Life Technologies) and 50 μg/ml gentamicin (Biochrom AG).

    Techniques: De-Phosphorylation Assay, Western Blot, Immunofluorescence, Infection

    VP6 in RV-infected cells. Shown is confocal immunofluorescence of MA104 cells infected with either OSU or SA11 (MOI, 25 VFU/cell) and transfected with siRNAs specific for SA11 VP6 or OSU VP6 or with a nontargeting siRNA (siNT) (A) or treated with 10 μM ML or DMSO (B). At the indicated times postinfection, viroplasms were visualized with anti-NSP5 antibody (red) and VP6 with MAb 4B2D2 (green). Scale bars, 5 μm.

    Journal: Journal of Virology

    Article Title: Identification of a Small Molecule That Compromises the Structural Integrity of Viroplasms and Rotavirus Double-Layered Particles

    doi: 10.1128/JVI.01943-17

    Figure Lengend Snippet: VP6 in RV-infected cells. Shown is confocal immunofluorescence of MA104 cells infected with either OSU or SA11 (MOI, 25 VFU/cell) and transfected with siRNAs specific for SA11 VP6 or OSU VP6 or with a nontargeting siRNA (siNT) (A) or treated with 10 μM ML or DMSO (B). At the indicated times postinfection, viroplasms were visualized with anti-NSP5 antibody (red) and VP6 with MAb 4B2D2 (green). Scale bars, 5 μm.

    Article Snippet: MA104 cells (embryonic African green monkey kidney cells; ATCC CRL-2378) were grown in Dulbecco's modified Eagle's medium (DMEM) (Life Technologies) containing 10% fetal bovine serum (Life Technologies) and 50 μg/ml gentamicin (Biochrom AG).

    Techniques: Infection, Immunofluorescence, Transfection

    Effect of ML on RV VP6. (A) VP6-VP2 interaction. Shown is Western blot analysis with anti-VP2 and anti-VP6 antibodies of immunoprecipitates (IP) obtained with anti-VP6 MAb RV138 from extracts of MA104 cells transfected with VP6 and VP2 and treated for 5 h with 10 μM ML or DMSO. The inhibitor (200 μM) was maintained during cell lysis and incubation with the precipitating antibody. The numbers on the left are kilodaltons. (B) VP6 trimer stability. (Left) Western blot analysis with anti-VP6 antibody of nonboiled extracts from MA104 cells infected with a recombinant vaccinia virus expressing VP6 (VVVP6) and treated with 10 μM ML or DMSO from 1 to 7 hpi. (Right) Western blot analysis of nonboiled extracts from cells infected with OSU (MOI, 25 VFU/cell) and treated with 10 μM ML or DMSO from 1 to 5 hpi. The numbers on the right are kilodaltons. (C) Confocal immunofluorescence (IF) analysis with the anti-VP6 MAb 4B2D2 of MA104 cells overexpressing VP6 (infected with VVVP6) and treated with 10 μM ML or DMSO from 1 to 7 hpi. The arrow indicates a VP6 higher-order structure observed in the absence of other RV proteins. Scale bars, 5 μm. (D) Representative images of VP6 tubes and spheres visualized by negative-staining electron microscopy after treatment with 25 μM ML for 4 h at 37°C. (E) Interaction of VP6 with ML evaluated by nanoscale thermophoresis. The fraction of Cys- or Lys-labeled VP6 bound to ML was plotted against increasing concentrations of the inhibitor. The data were fitted with two state equations, and an EC 50 of 294 ± 62 μM was calculated as the average of the results of three independent measurements. The error bars indicate standard deviations.

    Journal: Journal of Virology

    Article Title: Identification of a Small Molecule That Compromises the Structural Integrity of Viroplasms and Rotavirus Double-Layered Particles

    doi: 10.1128/JVI.01943-17

    Figure Lengend Snippet: Effect of ML on RV VP6. (A) VP6-VP2 interaction. Shown is Western blot analysis with anti-VP2 and anti-VP6 antibodies of immunoprecipitates (IP) obtained with anti-VP6 MAb RV138 from extracts of MA104 cells transfected with VP6 and VP2 and treated for 5 h with 10 μM ML or DMSO. The inhibitor (200 μM) was maintained during cell lysis and incubation with the precipitating antibody. The numbers on the left are kilodaltons. (B) VP6 trimer stability. (Left) Western blot analysis with anti-VP6 antibody of nonboiled extracts from MA104 cells infected with a recombinant vaccinia virus expressing VP6 (VVVP6) and treated with 10 μM ML or DMSO from 1 to 7 hpi. (Right) Western blot analysis of nonboiled extracts from cells infected with OSU (MOI, 25 VFU/cell) and treated with 10 μM ML or DMSO from 1 to 5 hpi. The numbers on the right are kilodaltons. (C) Confocal immunofluorescence (IF) analysis with the anti-VP6 MAb 4B2D2 of MA104 cells overexpressing VP6 (infected with VVVP6) and treated with 10 μM ML or DMSO from 1 to 7 hpi. The arrow indicates a VP6 higher-order structure observed in the absence of other RV proteins. Scale bars, 5 μm. (D) Representative images of VP6 tubes and spheres visualized by negative-staining electron microscopy after treatment with 25 μM ML for 4 h at 37°C. (E) Interaction of VP6 with ML evaluated by nanoscale thermophoresis. The fraction of Cys- or Lys-labeled VP6 bound to ML was plotted against increasing concentrations of the inhibitor. The data were fitted with two state equations, and an EC 50 of 294 ± 62 μM was calculated as the average of the results of three independent measurements. The error bars indicate standard deviations.

    Article Snippet: MA104 cells (embryonic African green monkey kidney cells; ATCC CRL-2378) were grown in Dulbecco's modified Eagle's medium (DMEM) (Life Technologies) containing 10% fetal bovine serum (Life Technologies) and 50 μg/ml gentamicin (Biochrom AG).

    Techniques: Western Blot, Transfection, Lysis, Incubation, Infection, Recombinant, Expressing, Immunofluorescence, Negative Staining, Electron Microscopy, Labeling

    ML antiviral activity is independent of cellular transcription and protein synthesis. (A) Western blot and confocal immunofluorescence analyses with the indicated antibodies of OSU-infected MA104 cells (25 VFU/cell) fed with EU and treated with 10 μM ML and/or 10 μg/ml actinomycin D (Act D) or DMSO for the indicated times. EU-labeled, newly synthesized RNAs were visualized by reaction with an Alexa-488-conjugated azide (green). Scale bars, 5 μm. (B) Western blot and confocal immunofluorescence analyses with the indicated antibodies of OSU-infected (MOI, 25 VFU/cell) MA104 cells treated with 10 μM ML and/or 10 μg/ml CHX or DMSO for the indicated times. Scale bars, 10 μm.

    Journal: Journal of Virology

    Article Title: Identification of a Small Molecule That Compromises the Structural Integrity of Viroplasms and Rotavirus Double-Layered Particles

    doi: 10.1128/JVI.01943-17

    Figure Lengend Snippet: ML antiviral activity is independent of cellular transcription and protein synthesis. (A) Western blot and confocal immunofluorescence analyses with the indicated antibodies of OSU-infected MA104 cells (25 VFU/cell) fed with EU and treated with 10 μM ML and/or 10 μg/ml actinomycin D (Act D) or DMSO for the indicated times. EU-labeled, newly synthesized RNAs were visualized by reaction with an Alexa-488-conjugated azide (green). Scale bars, 5 μm. (B) Western blot and confocal immunofluorescence analyses with the indicated antibodies of OSU-infected (MOI, 25 VFU/cell) MA104 cells treated with 10 μM ML and/or 10 μg/ml CHX or DMSO for the indicated times. Scale bars, 10 μm.

    Article Snippet: MA104 cells (embryonic African green monkey kidney cells; ATCC CRL-2378) were grown in Dulbecco's modified Eagle's medium (DMEM) (Life Technologies) containing 10% fetal bovine serum (Life Technologies) and 50 μg/ml gentamicin (Biochrom AG).

    Techniques: Activity Assay, Western Blot, Immunofluorescence, Infection, Activated Clotting Time Assay, Labeling, Synthesized

    ML effect on RV replication. (A to E) Western blot and confocal immunofluorescence analyses with the indicated antibodies of RV-infected cells (MOI, 25 VFU/cell) treated with ML at 10 μM, unless otherwise indicated, or with DMSO (D) for the indicated times. Scale bars, 10 μm. TUB, tubulin. (F) Time course of viral progeny yield of OSU-infected (MOI, 25 VFU/cell) MA104 cells treated with 10 μM ML, added at 2 hpi. The data are presented as averages ± standard deviations of the results of three independent experiments. ***, P

    Journal: Journal of Virology

    Article Title: Identification of a Small Molecule That Compromises the Structural Integrity of Viroplasms and Rotavirus Double-Layered Particles

    doi: 10.1128/JVI.01943-17

    Figure Lengend Snippet: ML effect on RV replication. (A to E) Western blot and confocal immunofluorescence analyses with the indicated antibodies of RV-infected cells (MOI, 25 VFU/cell) treated with ML at 10 μM, unless otherwise indicated, or with DMSO (D) for the indicated times. Scale bars, 10 μm. TUB, tubulin. (F) Time course of viral progeny yield of OSU-infected (MOI, 25 VFU/cell) MA104 cells treated with 10 μM ML, added at 2 hpi. The data are presented as averages ± standard deviations of the results of three independent experiments. ***, P

    Article Snippet: MA104 cells (embryonic African green monkey kidney cells; ATCC CRL-2378) were grown in Dulbecco's modified Eagle's medium (DMEM) (Life Technologies) containing 10% fetal bovine serum (Life Technologies) and 50 μg/ml gentamicin (Biochrom AG).

    Techniques: Western Blot, Immunofluorescence, Infection

    ML effect on VLS. Confocal immunofluorescence assay of VLS (A and C) and Western blot analysis (B) with the indicated antibodies of MA104 cells transfected with NSP5, NSP2, VP2, and VP6, as indicated. (A) NSP5 is shown in green and NSP2 or VP2 in red. (C) NSP5 is shown in red and VP6 in green. Cells were treated for 5 h with 10 μM ML or DMSO at 18 h posttransfection. Bars, 5 μm.

    Journal: Journal of Virology

    Article Title: Identification of a Small Molecule That Compromises the Structural Integrity of Viroplasms and Rotavirus Double-Layered Particles

    doi: 10.1128/JVI.01943-17

    Figure Lengend Snippet: ML effect on VLS. Confocal immunofluorescence assay of VLS (A and C) and Western blot analysis (B) with the indicated antibodies of MA104 cells transfected with NSP5, NSP2, VP2, and VP6, as indicated. (A) NSP5 is shown in green and NSP2 or VP2 in red. (C) NSP5 is shown in red and VP6 in green. Cells were treated for 5 h with 10 μM ML or DMSO at 18 h posttransfection. Bars, 5 μm.

    Article Snippet: MA104 cells (embryonic African green monkey kidney cells; ATCC CRL-2378) were grown in Dulbecco's modified Eagle's medium (DMEM) (Life Technologies) containing 10% fetal bovine serum (Life Technologies) and 50 μg/ml gentamicin (Biochrom AG).

    Techniques: Immunofluorescence, Western Blot, Transfection

    M s self-assembles. M i and M s were incubated at 500 nM or with equivalent amounts (monomer equivalent) of dimer and trimer for various times prior to resolution by SEC. Assemblies were monitored by reading the absorbance of fractions using micro BCA assay. ( A ) M i showed no self-association. ( B ) M s exhibited self-association over time. ( C,D ) Dimer and trimer were stable over time. ( E,F ) M i does not react with dimer or trimer to form larger assemblies. ( G,H ) M s reacts with dimer and trimer to form larger assemblies. See Figure 5—source data 1 . 10.7554/eLife.36584.012 Data for M s self-assembly.

    Journal: eLife

    Article Title: Inert and seed-competent tau monomers suggest structural origins of aggregation

    doi: 10.7554/eLife.36584

    Figure Lengend Snippet: M s self-assembles. M i and M s were incubated at 500 nM or with equivalent amounts (monomer equivalent) of dimer and trimer for various times prior to resolution by SEC. Assemblies were monitored by reading the absorbance of fractions using micro BCA assay. ( A ) M i showed no self-association. ( B ) M s exhibited self-association over time. ( C,D ) Dimer and trimer were stable over time. ( E,F ) M i does not react with dimer or trimer to form larger assemblies. ( G,H ) M s reacts with dimer and trimer to form larger assemblies. See Figure 5—source data 1 . 10.7554/eLife.36584.012 Data for M s self-assembly.

    Article Snippet: SEC fractions were frozen at −80°C after evaluation of protein content by Micro BCA assay (Thermo Scientific).

    Techniques: Incubation, Size-exclusion Chromatography, BIA-KA

    Pharmacological and gene silencing of autophagy markers blocks cocaine-mediated mitophagy. (a–f) Representative western blots showing expression of mitophagy markers such as PINK1 (a), PRKN (b), and DNM1L (c) and autophagy markers such as BECN1 (d), MAP1LC3B-II (e), and SQSTM1 (f) in mPMs pretreated with either 5 mM of 3-methyladenine (3-MA) or 100 nM of wortmannin for 1 h following exposure of cells to 10 μM cocaine for 24 h. (g–l) Representative western blots showing the expression of PINK1 (g), PRKN (h), and DNM1L (i), BECN1 (j), MAP1LC3B-II (k), and SQSTM1 (l) in mPMs transfected with BECN1 siRNA and scrambled siRNA following exposure of cells to 10 μM cocaine for 24 h. ACTB was probed as a protein loading control for all experiments. The data are presented as mean±SEM from six independent experiments. Non-parametric Kruskal-Wallis One-way ANOVA followed by Dunn’s post hoc test was used to determine the statistical significance among multiple groups. *, P

    Journal: Autophagy

    Article Title: Mitigation of cocaine-mediated mitochondrial damage, defective mitophagy and microglial activation by superoxide dismutase mimetics

    doi: 10.1080/15548627.2019.1607686

    Figure Lengend Snippet: Pharmacological and gene silencing of autophagy markers blocks cocaine-mediated mitophagy. (a–f) Representative western blots showing expression of mitophagy markers such as PINK1 (a), PRKN (b), and DNM1L (c) and autophagy markers such as BECN1 (d), MAP1LC3B-II (e), and SQSTM1 (f) in mPMs pretreated with either 5 mM of 3-methyladenine (3-MA) or 100 nM of wortmannin for 1 h following exposure of cells to 10 μM cocaine for 24 h. (g–l) Representative western blots showing the expression of PINK1 (g), PRKN (h), and DNM1L (i), BECN1 (j), MAP1LC3B-II (k), and SQSTM1 (l) in mPMs transfected with BECN1 siRNA and scrambled siRNA following exposure of cells to 10 μM cocaine for 24 h. ACTB was probed as a protein loading control for all experiments. The data are presented as mean±SEM from six independent experiments. Non-parametric Kruskal-Wallis One-way ANOVA followed by Dunn’s post hoc test was used to determine the statistical significance among multiple groups. *, P

    Article Snippet: Reagents Cocaine hydrochloride (C5776), 3-MA (M9281), wortmannin (W3144), rapamycin (R0395), bafilomycin A1 (B1793), Mdivi-1 (M0199), rotenone (557368), MitoTEMPO (SML0737) were purchased from Sigma-Aldrich.

    Techniques: Western Blot, Expressing, Transfection

    Effect of curcumin treatment on the cell cycle and apoptosis of NSCs in suspended state. NSCs were treated with control, curcumin and 3MA for 72 h. β-actin was used as the loading control. The relative level of protein was quantified by ImageJ. Experiments were repeated three times. NSC, neural stem cells; 3MA, 3-methyladenine.

    Journal: International Journal of Molecular Medicine

    Article Title: The effect of curcumin on the differentiation, apoptosis and cell cycle of neural stem cells is mediated through inhibiting autophagy by the modulation of Atg7 and p62

    doi: 10.3892/ijmm.2018.3847

    Figure Lengend Snippet: Effect of curcumin treatment on the cell cycle and apoptosis of NSCs in suspended state. NSCs were treated with control, curcumin and 3MA for 72 h. β-actin was used as the loading control. The relative level of protein was quantified by ImageJ. Experiments were repeated three times. NSC, neural stem cells; 3MA, 3-methyladenine.

    Article Snippet: 3-Methyladenine (3MA, an inhibitor of phosphatidylinositol 3-kinases and autophagosome formation; cat. no. s2767) was purchased from Selleck Chemicals (Shanghai, China).

    Techniques:

    Ultrastructural features of autophagy in NSCs treated with curcumin or 3MA for 72 h. (A) Negative control (untreated) cells; (B) NSCs treated with 10 µ M curcumin; (C) NSCs treated with 10 µ M 3MA, as a positive control. The autophagic vacuoles are indicated by arrowheads. Scale bar, 0.2 µ M; magnification, ×30,000. NSC, neural stem cells; 3MA, 3-methyladenine.

    Journal: International Journal of Molecular Medicine

    Article Title: The effect of curcumin on the differentiation, apoptosis and cell cycle of neural stem cells is mediated through inhibiting autophagy by the modulation of Atg7 and p62

    doi: 10.3892/ijmm.2018.3847

    Figure Lengend Snippet: Ultrastructural features of autophagy in NSCs treated with curcumin or 3MA for 72 h. (A) Negative control (untreated) cells; (B) NSCs treated with 10 µ M curcumin; (C) NSCs treated with 10 µ M 3MA, as a positive control. The autophagic vacuoles are indicated by arrowheads. Scale bar, 0.2 µ M; magnification, ×30,000. NSC, neural stem cells; 3MA, 3-methyladenine.

    Article Snippet: 3-Methyladenine (3MA, an inhibitor of phosphatidylinositol 3-kinases and autophagosome formation; cat. no. s2767) was purchased from Selleck Chemicals (Shanghai, China).

    Techniques: Negative Control, Positive Control

    Modulation of renal IRI by PHC, Rapa and a combination of 3-MA and PHC. (A) Hematoxylin eosin staining of renal tubular injury in the IRI model rats following treatment with PHC, 3-MA or Rapa (magnifications, ×200 or ×400). (B) Masson's trichrome staining of collagenous fibers in the IRI model rats following treatment with PHC, 3-MA or Rapa (magnifications, ×200 or ×400). (C) Periodic acid-Schiff staining of mesangial stromal and basement membrane thickening in renal tissues of the IRI model rats following treatment with PHC, 3-MA or Rapa (magnifications, ×200 or ×400). Pathogenic alterations are indicated by black arrows. All scale bars, 50 or 100 µm. 3-MA, 3-methyladenine; I/R, ischemia/reperfusion; IRI, I/R injury; PHC, penehyclidine hydrochloride; Rapa, rapamycin.

    Journal: Molecular Medicine Reports

    Article Title: Prevention of renal ischemia and reperfusion injury by penehyclidine hydrochloride through autophagy activation

    doi: 10.3892/mmr.2020.11024

    Figure Lengend Snippet: Modulation of renal IRI by PHC, Rapa and a combination of 3-MA and PHC. (A) Hematoxylin eosin staining of renal tubular injury in the IRI model rats following treatment with PHC, 3-MA or Rapa (magnifications, ×200 or ×400). (B) Masson's trichrome staining of collagenous fibers in the IRI model rats following treatment with PHC, 3-MA or Rapa (magnifications, ×200 or ×400). (C) Periodic acid-Schiff staining of mesangial stromal and basement membrane thickening in renal tissues of the IRI model rats following treatment with PHC, 3-MA or Rapa (magnifications, ×200 or ×400). Pathogenic alterations are indicated by black arrows. All scale bars, 50 or 100 µm. 3-MA, 3-methyladenine; I/R, ischemia/reperfusion; IRI, I/R injury; PHC, penehyclidine hydrochloride; Rapa, rapamycin.

    Article Snippet: PHC (0.45 mg/kg; Chengdu List Pharmaceutical, Co., Ltd.), 3-MA (0.5 mg/100 g; cat. no. S2767; Selleck Chemicals) and Rapa (1 mg/100 g; cat. no. AY22989; MedChemExpress) were intravenously administered to rats 30 min prior to I/R induction, as previously described ( ).

    Techniques: Staining

    Preparation and characterization of 2D net -poly(MA-11-2-11-MA). ( A ) Schematic illustration of the molecular structure of gemini monomer MA-11-2-11-MA and 2D net- poly(MA-11-2-11-MA). ( B ) AFM image and height analysis of the 2D net- poly(MA-11-2-11-MA) on the silicon substrate. Rq, root mean square roughness. ( C ) Simulation of the bilayer thickness with vertically arranged MA-11-2-11-MA monomers using Materials Studio. ( D ) TEM image of the freestanding film of 2D net -poly(MA-11-2-11-MA).

    Journal: Science Advances

    Article Title: Two-dimensional polymers with versatile functionalities via gemini monomers

    doi: 10.1126/sciadv.aaw9120

    Figure Lengend Snippet: Preparation and characterization of 2D net -poly(MA-11-2-11-MA). ( A ) Schematic illustration of the molecular structure of gemini monomer MA-11-2-11-MA and 2D net- poly(MA-11-2-11-MA). ( B ) AFM image and height analysis of the 2D net- poly(MA-11-2-11-MA) on the silicon substrate. Rq, root mean square roughness. ( C ) Simulation of the bilayer thickness with vertically arranged MA-11-2-11-MA monomers using Materials Studio. ( D ) TEM image of the freestanding film of 2D net -poly(MA-11-2-11-MA).

    Article Snippet: The conversion ratio of MA-11-2-11-MA was close to 100%, which was important for the formation of a highly cross-linked 2D network (fig. S2, A and B).

    Techniques: Transmission Electron Microscopy

    DPD simulation of 2D net -poly(MA-11-2-11-MA). ( A ) Schematic illustration of the coarse-grained model of MA-11-2-11-MA. ( B ) Left: A representative snapshot of the MA-11-2-11-MA bilayer before polymerization. The conversion ratio of methacrylate groups is 0%. Right: Density profile of each component distributed in the vertical direction. Alkyl segments (black), ammonium groups (blue), methacrylate groups (red), and water (yellow). ( C ) Snapshots of MA-11-2-11-MA bilayers at increasing conversion ratios of methacrylate groups. ( D ) Left: A representative snapshot of the net -poly(MA-11-2-11-MA) bilayer after polymerization. The inset represents a top view of the poly(methacrylate) sublayer. The conversion ratio of the methacrylate groups is 98%. Right: Density profile of each component distributed in the vertical direction.

    Journal: Science Advances

    Article Title: Two-dimensional polymers with versatile functionalities via gemini monomers

    doi: 10.1126/sciadv.aaw9120

    Figure Lengend Snippet: DPD simulation of 2D net -poly(MA-11-2-11-MA). ( A ) Schematic illustration of the coarse-grained model of MA-11-2-11-MA. ( B ) Left: A representative snapshot of the MA-11-2-11-MA bilayer before polymerization. The conversion ratio of methacrylate groups is 0%. Right: Density profile of each component distributed in the vertical direction. Alkyl segments (black), ammonium groups (blue), methacrylate groups (red), and water (yellow). ( C ) Snapshots of MA-11-2-11-MA bilayers at increasing conversion ratios of methacrylate groups. ( D ) Left: A representative snapshot of the net -poly(MA-11-2-11-MA) bilayer after polymerization. The inset represents a top view of the poly(methacrylate) sublayer. The conversion ratio of the methacrylate groups is 98%. Right: Density profile of each component distributed in the vertical direction.

    Article Snippet: The conversion ratio of MA-11-2-11-MA was close to 100%, which was important for the formation of a highly cross-linked 2D network (fig. S2, A and B).

    Techniques:

    NMR characterization of MA-11-2-11-MA assembly before and after polymerization. ( A ) Schematic illustration of the proton number in MA-11-2-11-MA. ( B ) 1 H NMR spectra of MA-11-2-11-MA assembly in D 2 O at 65°C (top) and the corresponding ROESY spectra by the selective irradiation of proton H-9 (bottom). ( C ) ROESY spectra of MA-11-2-11-MA assembly by the selective irradiation of H-9 at increased conversion ratios of methacrylate groups. The conversion ratios were estimated from the FT-IR spectra.

    Journal: Science Advances

    Article Title: Two-dimensional polymers with versatile functionalities via gemini monomers

    doi: 10.1126/sciadv.aaw9120

    Figure Lengend Snippet: NMR characterization of MA-11-2-11-MA assembly before and after polymerization. ( A ) Schematic illustration of the proton number in MA-11-2-11-MA. ( B ) 1 H NMR spectra of MA-11-2-11-MA assembly in D 2 O at 65°C (top) and the corresponding ROESY spectra by the selective irradiation of proton H-9 (bottom). ( C ) ROESY spectra of MA-11-2-11-MA assembly by the selective irradiation of H-9 at increased conversion ratios of methacrylate groups. The conversion ratios were estimated from the FT-IR spectra.

    Article Snippet: The conversion ratio of MA-11-2-11-MA was close to 100%, which was important for the formation of a highly cross-linked 2D network (fig. S2, A and B).

    Techniques: Nuclear Magnetic Resonance, Irradiation

    Functionalization of 2D net -poly(MA-11-2-11-MA). ( A ) Chemical structures of the monomeric derivatives (MA-11-F; F, various functional groups) and copolymerization with MA-11-2-11-MA. ( B ) Fluorescence films of net- poly(MA-11-2-11-MA)- co -(MA-11-F) (F = RhB, Fls, or Pyr) in number and letter shapes (scale bar, 500 μm). ( C ) Fluorescence microscope images of net- poly(MA-11-2-11-MA)- co -(MA-11-C≡C)–coated silica wool after post-polymerization functionalization with RhB-EO 4 -N 3 , Fls-EO 4 -N 3 , and Pyr-EO 4 -N 3 via click reactions (scale bars, 200 μm).

    Journal: Science Advances

    Article Title: Two-dimensional polymers with versatile functionalities via gemini monomers

    doi: 10.1126/sciadv.aaw9120

    Figure Lengend Snippet: Functionalization of 2D net -poly(MA-11-2-11-MA). ( A ) Chemical structures of the monomeric derivatives (MA-11-F; F, various functional groups) and copolymerization with MA-11-2-11-MA. ( B ) Fluorescence films of net- poly(MA-11-2-11-MA)- co -(MA-11-F) (F = RhB, Fls, or Pyr) in number and letter shapes (scale bar, 500 μm). ( C ) Fluorescence microscope images of net- poly(MA-11-2-11-MA)- co -(MA-11-C≡C)–coated silica wool after post-polymerization functionalization with RhB-EO 4 -N 3 , Fls-EO 4 -N 3 , and Pyr-EO 4 -N 3 via click reactions (scale bars, 200 μm).

    Article Snippet: The conversion ratio of MA-11-2-11-MA was close to 100%, which was important for the formation of a highly cross-linked 2D network (fig. S2, A and B).

    Techniques: Functional Assay, Fluorescence, Microscopy

    TCP8, TCP14, and TCP15 interact with NPR1. (A) Interactions between NPR1 with TCP8, TCP14, and TCP15 in yeast two-hybrid assays. Diploid yeast cells were serially diluted at OD 600 = 1.0, 0.1, and 0.01. Aliquots of 10 μL of each dilution were plated on double synthetic dropout control plates without leucine and tryptophan (-LW) and selective triple synthetic dropout plates without leucine, tryptophan, and histidine with 1 mM 3-amino-1,2,4-triazole (–LWH + 1 mM 3AT). Photographs were taken 5 days after plating. EV, Empty vector; BD, GAL4 DNA-binding domain; AD, GAL4 activation domain. (B) Co-immunoprecipitation (Co-IP) of NPR1 with TCP8 in Nicotiana benthamiana . NPR1-GFP and FLAG-TCP8 under control of the cauliflower mosaic virus (CMV) 35S promoter were transiently co-expressed in N . benthamiana by agroinfiltration. Total protein extracts were immunoprecipitated with anti-FLAG M2 magnetic beads. The input and immunoprecipitated protein were analyzed by immunoblot using the anti-FLAG and anti-GFP antibodies. kDa, kilodaltons. (C,D) Co-IP of TCP14 and TCP15 with NPR1 in N . benthamiana . Constitutively expressed NPR1-GFP was transiently co-expressed with HA-TCP14 (C) and HA-TCP15 (D) under control of the CMV 35S promoter in N . benthamiana by agroinfiltration. Total protein extracts were immunoprecipitated using anti-GFP trap beads. The input and immuno-precipitated proteins were analyzed by immunoblot using anti-HA and anti-GFP antibodies.

    Journal: Frontiers in Plant Science

    Article Title: TCP Transcription Factors Interact With NPR1 and Contribute Redundantly to Systemic Acquired Resistance

    doi: 10.3389/fpls.2018.01153

    Figure Lengend Snippet: TCP8, TCP14, and TCP15 interact with NPR1. (A) Interactions between NPR1 with TCP8, TCP14, and TCP15 in yeast two-hybrid assays. Diploid yeast cells were serially diluted at OD 600 = 1.0, 0.1, and 0.01. Aliquots of 10 μL of each dilution were plated on double synthetic dropout control plates without leucine and tryptophan (-LW) and selective triple synthetic dropout plates without leucine, tryptophan, and histidine with 1 mM 3-amino-1,2,4-triazole (–LWH + 1 mM 3AT). Photographs were taken 5 days after plating. EV, Empty vector; BD, GAL4 DNA-binding domain; AD, GAL4 activation domain. (B) Co-immunoprecipitation (Co-IP) of NPR1 with TCP8 in Nicotiana benthamiana . NPR1-GFP and FLAG-TCP8 under control of the cauliflower mosaic virus (CMV) 35S promoter were transiently co-expressed in N . benthamiana by agroinfiltration. Total protein extracts were immunoprecipitated with anti-FLAG M2 magnetic beads. The input and immunoprecipitated protein were analyzed by immunoblot using the anti-FLAG and anti-GFP antibodies. kDa, kilodaltons. (C,D) Co-IP of TCP14 and TCP15 with NPR1 in N . benthamiana . Constitutively expressed NPR1-GFP was transiently co-expressed with HA-TCP14 (C) and HA-TCP15 (D) under control of the CMV 35S promoter in N . benthamiana by agroinfiltration. Total protein extracts were immunoprecipitated using anti-GFP trap beads. The input and immuno-precipitated proteins were analyzed by immunoblot using anti-HA and anti-GFP antibodies.

    Article Snippet: For the interaction of NPR1 with TCP14 or TCP15, protein extracts were incubated with 20 μl GFP-Trap®_ MA Beads (Chromotek) with gentle rotation at 4°C overnight.

    Techniques: Plasmid Preparation, Binding Assay, Activation Assay, Immunoprecipitation, Co-Immunoprecipitation Assay, Magnetic Beads

    TCP15 binds to the PR5 promoter at the TCP binding site. (A) Interactions between the TCP transcription factors and the PR promoters in yeast one-hybrid system (Y1H). Constructs expressing pDEST22-TCPs and empty pDEST22 (EV) were transformed into the yeast strain YU. The PR promoters fused with LacZ reporter gene were integrated into the yeast strain YM4271. Healthy diploids grew on dropout medium without tryptophan and uracil were selected. Binding ability between TCPs and the PR promoters was quantified by β-galactosidase activity which was calculated in fold change over EV control. Three-fold induction was set as the cut-off. The binding ability of TCP8, TCP14, and TCP15 to the PR1 promoter (upper panel), the PR2 promoter (middle panel), and the PR5 promoter (lower panel) is shown. Error bars represent SD of three repeats. (B) Diagram of the PR5 gene structure. (+1) means transcription start site; long line indicates the promoter of PR5 ; short lines marked by a, b, and c represent the fragments amplified by RT-PCR in (D) ; star shows TCP binding site located at −788 bp; TCPm represents TCP binding site with mutations. (C) Interactions between TCP15 with the PR5 promoter containing a TCPm and the PR5 promoter (500 bp) in Y1H assays. Error bars represent SD of three repeats. (D) Association between TCP15 and the PR5 promoter was studied by chromatin immunoprecipitation (ChIP) assays combined with RT-PCR analysis. Twelve-day-old seedlings of pTA:AtTCP15-EYFP and 35S:GFP transgenic plants were treated with 30 μM DEX for 24 h. Samples were collected at another 24 h after 0.5 mM SA induction. The enrichment of PR5 promoter DNA in immunoprecipitated samples was shown as % input. Error bars represent SD of three technical repeats. Statistical analysis was studied by t -test (* p

    Journal: Frontiers in Plant Science

    Article Title: TCP Transcription Factors Interact With NPR1 and Contribute Redundantly to Systemic Acquired Resistance

    doi: 10.3389/fpls.2018.01153

    Figure Lengend Snippet: TCP15 binds to the PR5 promoter at the TCP binding site. (A) Interactions between the TCP transcription factors and the PR promoters in yeast one-hybrid system (Y1H). Constructs expressing pDEST22-TCPs and empty pDEST22 (EV) were transformed into the yeast strain YU. The PR promoters fused with LacZ reporter gene were integrated into the yeast strain YM4271. Healthy diploids grew on dropout medium without tryptophan and uracil were selected. Binding ability between TCPs and the PR promoters was quantified by β-galactosidase activity which was calculated in fold change over EV control. Three-fold induction was set as the cut-off. The binding ability of TCP8, TCP14, and TCP15 to the PR1 promoter (upper panel), the PR2 promoter (middle panel), and the PR5 promoter (lower panel) is shown. Error bars represent SD of three repeats. (B) Diagram of the PR5 gene structure. (+1) means transcription start site; long line indicates the promoter of PR5 ; short lines marked by a, b, and c represent the fragments amplified by RT-PCR in (D) ; star shows TCP binding site located at −788 bp; TCPm represents TCP binding site with mutations. (C) Interactions between TCP15 with the PR5 promoter containing a TCPm and the PR5 promoter (500 bp) in Y1H assays. Error bars represent SD of three repeats. (D) Association between TCP15 and the PR5 promoter was studied by chromatin immunoprecipitation (ChIP) assays combined with RT-PCR analysis. Twelve-day-old seedlings of pTA:AtTCP15-EYFP and 35S:GFP transgenic plants were treated with 30 μM DEX for 24 h. Samples were collected at another 24 h after 0.5 mM SA induction. The enrichment of PR5 promoter DNA in immunoprecipitated samples was shown as % input. Error bars represent SD of three technical repeats. Statistical analysis was studied by t -test (* p

    Article Snippet: For the interaction of NPR1 with TCP14 or TCP15, protein extracts were incubated with 20 μl GFP-Trap®_ MA Beads (Chromotek) with gentle rotation at 4°C overnight.

    Techniques: Binding Assay, Construct, Expressing, Transformation Assay, Activity Assay, Amplification, Reverse Transcription Polymerase Chain Reaction, Chromatin Immunoprecipitation, Transgenic Assay, Immunoprecipitation

    INPP4B is upregulated by NPM1-mA in leukemia cells via ERK/Ets-1 signaling. ( a ) qRT-PCR analysis of INPP4B mRNA expression, ( b ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. ( c ) qRT-PCR analysis of INPP4B mRNA expression, ( d ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the THP-1 and K562 cells transduced with the plasmids expressing NPM1-wt or NPM1-mA. ( e ) qRT-PCR analysis of INPP4B mRNA expression, ( f ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3, p-Ets-1 and Ets-1 from the OCI-AML3 cells transfected with the control siRNA or siEts-1. ( g ) Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 and SGK3, ( h ) qRT-PCR analysis of INPP4B mRNA expression from the OCI-AML3 cells treated with different concentration of PD98059 (0, 10, 20 and 40 μM). i Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. Proteins were quantified using image software and normalized against β-actin. Data were represented as mean ± s.d. of three individual experiments. * p

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: INPP4B promotes cell survival via SGK3 activation in NPM1-mutated leukemia

    doi: 10.1186/s13046-018-0675-9

    Figure Lengend Snippet: INPP4B is upregulated by NPM1-mA in leukemia cells via ERK/Ets-1 signaling. ( a ) qRT-PCR analysis of INPP4B mRNA expression, ( b ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. ( c ) qRT-PCR analysis of INPP4B mRNA expression, ( d ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the THP-1 and K562 cells transduced with the plasmids expressing NPM1-wt or NPM1-mA. ( e ) qRT-PCR analysis of INPP4B mRNA expression, ( f ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3, p-Ets-1 and Ets-1 from the OCI-AML3 cells transfected with the control siRNA or siEts-1. ( g ) Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 and SGK3, ( h ) qRT-PCR analysis of INPP4B mRNA expression from the OCI-AML3 cells treated with different concentration of PD98059 (0, 10, 20 and 40 μM). i Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. Proteins were quantified using image software and normalized against β-actin. Data were represented as mean ± s.d. of three individual experiments. * p

    Article Snippet: Moreover, our previous study has verified that ERK signaling is continuously activated by NPM1-mA [ ].

    Techniques: Quantitative RT-PCR, Expressing, Western Blot, Transduction, Transfection, Concentration Assay, Software

    NPM1-mA-mediated INPP4B upregulation promotes cell proliferation in OCI-AML3 cells. The NPM1-mA-silenced OCI-AML3 cells were subjected to ( a ) CCK8 assays and ( b ) colony forming assays. c The NPM1-mA-silenced OCI-AML3 cells were transfected with the pEAK-Flag/INPP4B plasmids, western blotting analysis of INPP4B and NPM1-mA. Proteins were quantified using image software and normalized against β-actin. d CCK-8 assay analysis of cell proliferation in NPM1-mA-silenced OCI-AML3 cells, followed by Flag-INPP4B introduction. Data were represented as mean ± s.d. of three individual experiments. * p

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: INPP4B promotes cell survival via SGK3 activation in NPM1-mutated leukemia

    doi: 10.1186/s13046-018-0675-9

    Figure Lengend Snippet: NPM1-mA-mediated INPP4B upregulation promotes cell proliferation in OCI-AML3 cells. The NPM1-mA-silenced OCI-AML3 cells were subjected to ( a ) CCK8 assays and ( b ) colony forming assays. c The NPM1-mA-silenced OCI-AML3 cells were transfected with the pEAK-Flag/INPP4B plasmids, western blotting analysis of INPP4B and NPM1-mA. Proteins were quantified using image software and normalized against β-actin. d CCK-8 assay analysis of cell proliferation in NPM1-mA-silenced OCI-AML3 cells, followed by Flag-INPP4B introduction. Data were represented as mean ± s.d. of three individual experiments. * p

    Article Snippet: Moreover, our previous study has verified that ERK signaling is continuously activated by NPM1-mA [ ].

    Techniques: Transfection, Western Blot, Software, CCK-8 Assay

    High expression of INPP4B is associated with poor survival outcome in NPM1-mutated leukemia. Kaplan-Meier survival data of 153 AML patients were used to analysis ( a ) OS and ( b ) EFS curves according to INPP4B levels. c Heatmap of 38 primary AML samples with NPM1 mutation from TCGA dataset in which INPP4B expression was aligned with patient event (Survival or Death). Kaplan-Meier survival data of patients with NPM1-mutated AML were used to analysis ( d ) OS and ( e ) EFS curves according to INPP4B levels. f Schematic diagram describing the functional significance of INPP4B in the NPM1-mutated leukemia cells. INPP4B promotes leukemia cell survival in a SGK3-dependent and AKT-independent manner. The expression of INPP4B partially upregulated by NPM1-mA is due to ERK/Ets-1 signaling

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: INPP4B promotes cell survival via SGK3 activation in NPM1-mutated leukemia

    doi: 10.1186/s13046-018-0675-9

    Figure Lengend Snippet: High expression of INPP4B is associated with poor survival outcome in NPM1-mutated leukemia. Kaplan-Meier survival data of 153 AML patients were used to analysis ( a ) OS and ( b ) EFS curves according to INPP4B levels. c Heatmap of 38 primary AML samples with NPM1 mutation from TCGA dataset in which INPP4B expression was aligned with patient event (Survival or Death). Kaplan-Meier survival data of patients with NPM1-mutated AML were used to analysis ( d ) OS and ( e ) EFS curves according to INPP4B levels. f Schematic diagram describing the functional significance of INPP4B in the NPM1-mutated leukemia cells. INPP4B promotes leukemia cell survival in a SGK3-dependent and AKT-independent manner. The expression of INPP4B partially upregulated by NPM1-mA is due to ERK/Ets-1 signaling

    Article Snippet: Moreover, our previous study has verified that ERK signaling is continuously activated by NPM1-mA [ ].

    Techniques: Expressing, Mutagenesis, Functional Assay

    High expression levels of INPP4B in leukemia cells with the NPM1 mutation. a RNA-seq mRNA expression data from the TCGA database were used to compare INPP4B expression between AML patients with ( n = 41) and without NPM1 mutation ( n = 130). * p

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: INPP4B promotes cell survival via SGK3 activation in NPM1-mutated leukemia

    doi: 10.1186/s13046-018-0675-9

    Figure Lengend Snippet: High expression levels of INPP4B in leukemia cells with the NPM1 mutation. a RNA-seq mRNA expression data from the TCGA database were used to compare INPP4B expression between AML patients with ( n = 41) and without NPM1 mutation ( n = 130). * p

    Article Snippet: Moreover, our previous study has verified that ERK signaling is continuously activated by NPM1-mA [ ].

    Techniques: Expressing, Mutagenesis, RNA Sequencing Assay

    Effect of SAV7472-SAV7473 overexpression on avermectin production in S. avermitilis ATCC 31267. The strains (left to right) are wild-type ATCC 31267, plasmid control strain ATCC 31267(pKC1139), and SAV7472-SAV7473 overexpressing transformants 31267(pKCE7273)-1,

    Journal: Journal of Bacteriology

    Article Title: Characterization of SAV7471, a TetR-Family Transcriptional Regulator Involved in the Regulation of Coenzyme A Metabolism in Streptomyces avermitilis

    doi: 10.1128/JB.00716-13

    Figure Lengend Snippet: Effect of SAV7472-SAV7473 overexpression on avermectin production in S. avermitilis ATCC 31267. The strains (left to right) are wild-type ATCC 31267, plasmid control strain ATCC 31267(pKC1139), and SAV7472-SAV7473 overexpressing transformants 31267(pKCE7273)-1,

    Article Snippet: In this study, we describe the regulatory role of SAV7471 in avermectin production in S. avermitilis ATCC 31267.

    Techniques: Over Expression, Plasmid Preparation

    Effect of overexpression and deletion of the SAV7471 gene on avermectin production in S. avermitilis ATCC 31267. The strains left to right are wild-type ATCC 31267, the empty-plasmid-containing control ATCC 31267(pSET152), the SAV7471 -overexpressing transformants

    Journal: Journal of Bacteriology

    Article Title: Characterization of SAV7471, a TetR-Family Transcriptional Regulator Involved in the Regulation of Coenzyme A Metabolism in Streptomyces avermitilis

    doi: 10.1128/JB.00716-13

    Figure Lengend Snippet: Effect of overexpression and deletion of the SAV7471 gene on avermectin production in S. avermitilis ATCC 31267. The strains left to right are wild-type ATCC 31267, the empty-plasmid-containing control ATCC 31267(pSET152), the SAV7471 -overexpressing transformants

    Article Snippet: In this study, we describe the regulatory role of SAV7471 in avermectin production in S. avermitilis ATCC 31267.

    Techniques: Over Expression, Plasmid Preparation