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  • 99
    New England Biolabs cpg methyltransferase
    Cpg Methyltransferase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 780 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 780 article reviews
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    90
    Mimetics hiv 1 transcription
    Model of <t>HIV-1</t> transcriptional activation by Tat and RelB. HIV-1 Tat interacts with RelB. RelB and Tat are recruited to LTR DNA to assist transcription, which increases recruitment of RNA Pol II to the LTR and leads to greater expression of viral genes
    Hiv 1 Transcription, supplied by Mimetics, used in various techniques. Bioz Stars score: 90/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Novartis ectrims
    Model of <t>HIV-1</t> transcriptional activation by Tat and RelB. HIV-1 Tat interacts with RelB. RelB and Tat are recruited to LTR DNA to assist transcription, which increases recruitment of RNA Pol II to the LTR and leads to greater expression of viral genes
    Ectrims, supplied by Novartis, used in various techniques. Bioz Stars score: 89/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Brinkmann Instruments brinkmann ao
    Model of <t>HIV-1</t> transcriptional activation by Tat and RelB. HIV-1 Tat interacts with RelB. RelB and Tat are recruited to LTR DNA to assist transcription, which increases recruitment of RNA Pol II to the LTR and leads to greater expression of viral genes
    Brinkmann Ao, supplied by Brinkmann Instruments, used in various techniques. Bioz Stars score: 92/100, based on 1349 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Novartis kknms bundesministerium für bildung und forschung
    Model of <t>HIV-1</t> transcriptional activation by Tat and RelB. HIV-1 Tat interacts with RelB. RelB and Tat are recruited to LTR DNA to assist transcription, which increases recruitment of RNA Pol II to the LTR and leads to greater expression of viral genes
    Kknms Bundesministerium Für Bildung Und Forschung, supplied by Novartis, used in various techniques. Bioz Stars score: 88/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    ZeptoMetrix p19gag
    Inhibitory effect of IFN on Gag-raft association. IFN increases the detergent solubility of Gag (A). ACH-transfected 293T cells in the absence (upper panel) or presence (lower panel) of IFN were treated with or without 0.5% Triton X-100 (Tx100) on ice for 20 min before fixation. Gag membrane localization was detected by confocal analysis by using anti-p19 Gag . In the presence of Triton X-100, the punctate fluorescence pattern was largely preserved in DRMs without IFN. However, it was largely lost in DRMs from cells treated with 100 U of IFN/ml. IFN blocks Gag-raft colocalization (B). ACH-expressing cells were stained with anti-p19 Gag MAb (green) and anti-GM1 (red). p19 Gag immunofluorescence colocalized with raft-associated marker GM1 in the absence of IFN (the upper panel), whereas only a small portion of <t>p19Gag</t> was colocalized with GM1 following IFN treatment (the lower panel).
    P19gag, supplied by ZeptoMetrix, used in various techniques. Bioz Stars score: 88/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Qiagen polyfect reagent
    Inhibitory effect of IFN on Gag-raft association. IFN increases the detergent solubility of Gag (A). ACH-transfected 293T cells in the absence (upper panel) or presence (lower panel) of IFN were treated with or without 0.5% Triton X-100 (Tx100) on ice for 20 min before fixation. Gag membrane localization was detected by confocal analysis by using anti-p19 Gag . In the presence of Triton X-100, the punctate fluorescence pattern was largely preserved in DRMs without IFN. However, it was largely lost in DRMs from cells treated with 100 U of IFN/ml. IFN blocks Gag-raft colocalization (B). ACH-expressing cells were stained with anti-p19 Gag MAb (green) and anti-GM1 (red). p19 Gag immunofluorescence colocalized with raft-associated marker GM1 in the absence of IFN (the upper panel), whereas only a small portion of <t>p19Gag</t> was colocalized with GM1 following IFN treatment (the lower panel).
    Polyfect Reagent, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 2087 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Mimetics smac mimetics
    <t>SMAC</t> mimetics induce the degradation of BIRC2 and XIAP in <t>HIV-T</t> CM . ( A ) Left , representative western blots of FAS, membrane-bound FASLG (mFASLG) and soluble FASLG (sFASLG) in T CM and HIV-T CM . Right , densitometric analysis of blots. n = 4. ( B ) Left , representative western blots of FAS, membrane-bound FASLG (mFASLG) and soluble FASLG (sFASLG) in A3.01 and ACH-2. Experiment was performed in triplicate. Right , densitometric analysis of blots. ( C ) Left , representative western blots of XIAP and BIRC2 in A3.01 and ACH-2. Experiment was performed in triplicate. Right , densitometric analysis of blots. ( D ) Left , representative western blots of XIAP and BIRC2 in T CM and HIV-T CM using antibody to XIAP, BIRC2 and ACTB. Right , densitometric analysis of blots. n = 4. ( E ) Left , representative western blots of LC3B isoforms, BECN1 and SQSTM1 in T CM and HIV-T CM . Right , densitometric analysis of blots. n = 4. ( F ) HIV-T CM were treated for 4 h with 100 nM bafilomycin A 1 . Left , representative western blots of LC3B isoforms and SQSTM1. Right , densitometric analysis of blots. n = 4. ( G ) T CM and HIV-T CM were treated for 24 h with increasing concentrations of birinapant, GDC-0152, or embelin. Left , representative western blots of XIAP and BIRC2. Right , densitometric analysis of blots. n = 4.
    Smac Mimetics, supplied by Mimetics, used in various techniques. Bioz Stars score: 90/100, based on 299 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher control sirna
    ADAR1 proviral effect is mediated through inhibition of <t>PKR</t> activation. 293-T cells were transfected with 4 μg of (A) pACH or (B) pH6neo molecular clones and increasing amount (0, 5, 50, 500 ng) of ADAR1 plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-Tax1, anti-Tax2, anti-PKR, anti-phospho-PKR and anti-actin antibodies. (C, D) : 293-T cells were transfected with <t>siRNA</t> directed against PKR (20nM) or with control siRNA. Twelve hours later cells were transfected with 20nM of the same siRNA together with 1.2 μg of (C) pACH, (D) pH6neo and 125 ng of an ADAR1 expression plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-PKR, anti-phospho-PKR and anti-actin antibodies.
    Control Sirna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 15990 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Promega fugene hd
    ADAR1 proviral effect is mediated through inhibition of <t>PKR</t> activation. 293-T cells were transfected with 4 μg of (A) pACH or (B) pH6neo molecular clones and increasing amount (0, 5, 50, 500 ng) of ADAR1 plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-Tax1, anti-Tax2, anti-PKR, anti-phospho-PKR and anti-actin antibodies. (C, D) : 293-T cells were transfected with <t>siRNA</t> directed against PKR (20nM) or with control siRNA. Twelve hours later cells were transfected with 20nM of the same siRNA together with 1.2 μg of (C) pACH, (D) pH6neo and 125 ng of an ADAR1 expression plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-PKR, anti-phospho-PKR and anti-actin antibodies.
    Fugene Hd, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 15329 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher pcr 4 topo vector
    ADAR1 proviral effect is mediated through inhibition of <t>PKR</t> activation. 293-T cells were transfected with 4 μg of (A) pACH or (B) pH6neo molecular clones and increasing amount (0, 5, 50, 500 ng) of ADAR1 plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-Tax1, anti-Tax2, anti-PKR, anti-phospho-PKR and anti-actin antibodies. (C, D) : 293-T cells were transfected with <t>siRNA</t> directed against PKR (20nM) or with control siRNA. Twelve hours later cells were transfected with 20nM of the same siRNA together with 1.2 μg of (C) pACH, (D) pH6neo and 125 ng of an ADAR1 expression plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-PKR, anti-phospho-PKR and anti-actin antibodies.
    Pcr 4 Topo Vector, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3237 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Stratagene real time pcr
    ADAR1 proviral effect is mediated through inhibition of <t>PKR</t> activation. 293-T cells were transfected with 4 μg of (A) pACH or (B) pH6neo molecular clones and increasing amount (0, 5, 50, 500 ng) of ADAR1 plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-Tax1, anti-Tax2, anti-PKR, anti-phospho-PKR and anti-actin antibodies. (C, D) : 293-T cells were transfected with <t>siRNA</t> directed against PKR (20nM) or with control siRNA. Twelve hours later cells were transfected with 20nM of the same siRNA together with 1.2 μg of (C) pACH, (D) pH6neo and 125 ng of an ADAR1 expression plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-PKR, anti-phospho-PKR and anti-actin antibodies.
    Real Time Pcr, supplied by Stratagene, used in various techniques. Bioz Stars score: 92/100, based on 1824 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Qiagen hiperfect reagent
    ADAR1 proviral effect is mediated through inhibition of <t>PKR</t> activation. 293-T cells were transfected with 4 μg of (A) pACH or (B) pH6neo molecular clones and increasing amount (0, 5, 50, 500 ng) of ADAR1 plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-Tax1, anti-Tax2, anti-PKR, anti-phospho-PKR and anti-actin antibodies. (C, D) : 293-T cells were transfected with <t>siRNA</t> directed against PKR (20nM) or with control siRNA. Twelve hours later cells were transfected with 20nM of the same siRNA together with 1.2 μg of (C) pACH, (D) pH6neo and 125 ng of an ADAR1 expression plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-PKR, anti-phospho-PKR and anti-actin antibodies.
    Hiperfect Reagent, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 2258 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Brinkmann Instruments huikeshoven fj
    ADAR1 proviral effect is mediated through inhibition of <t>PKR</t> activation. 293-T cells were transfected with 4 μg of (A) pACH or (B) pH6neo molecular clones and increasing amount (0, 5, 50, 500 ng) of ADAR1 plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-Tax1, anti-Tax2, anti-PKR, anti-phospho-PKR and anti-actin antibodies. (C, D) : 293-T cells were transfected with <t>siRNA</t> directed against PKR (20nM) or with control siRNA. Twelve hours later cells were transfected with 20nM of the same siRNA together with 1.2 μg of (C) pACH, (D) pH6neo and 125 ng of an ADAR1 expression plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-PKR, anti-phospho-PKR and anti-actin antibodies.
    Huikeshoven Fj, supplied by Brinkmann Instruments, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Endovasc j endovasc ther
    ADAR1 proviral effect is mediated through inhibition of <t>PKR</t> activation. 293-T cells were transfected with 4 μg of (A) pACH or (B) pH6neo molecular clones and increasing amount (0, 5, 50, 500 ng) of ADAR1 plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-Tax1, anti-Tax2, anti-PKR, anti-phospho-PKR and anti-actin antibodies. (C, D) : 293-T cells were transfected with <t>siRNA</t> directed against PKR (20nM) or with control siRNA. Twelve hours later cells were transfected with 20nM of the same siRNA together with 1.2 μg of (C) pACH, (D) pH6neo and 125 ng of an ADAR1 expression plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-PKR, anti-phospho-PKR and anti-actin antibodies.
    J Endovasc Ther, supplied by Endovasc, used in various techniques. Bioz Stars score: 97/100, based on 2128 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher neon transfection system
    ADAR1 proviral effect is mediated through inhibition of <t>PKR</t> activation. 293-T cells were transfected with 4 μg of (A) pACH or (B) pH6neo molecular clones and increasing amount (0, 5, 50, 500 ng) of ADAR1 plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-Tax1, anti-Tax2, anti-PKR, anti-phospho-PKR and anti-actin antibodies. (C, D) : 293-T cells were transfected with <t>siRNA</t> directed against PKR (20nM) or with control siRNA. Twelve hours later cells were transfected with 20nM of the same siRNA together with 1.2 μg of (C) pACH, (D) pH6neo and 125 ng of an ADAR1 expression plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-PKR, anti-phospho-PKR and anti-actin antibodies.
    Neon Transfection System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 15041 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Stratagene quickchange site directed mutagenesis kit
    ADAR1 proviral effect is mediated through inhibition of <t>PKR</t> activation. 293-T cells were transfected with 4 μg of (A) pACH or (B) pH6neo molecular clones and increasing amount (0, 5, 50, 500 ng) of ADAR1 plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-Tax1, anti-Tax2, anti-PKR, anti-phospho-PKR and anti-actin antibodies. (C, D) : 293-T cells were transfected with <t>siRNA</t> directed against PKR (20nM) or with control siRNA. Twelve hours later cells were transfected with 20nM of the same siRNA together with 1.2 μg of (C) pACH, (D) pH6neo and 125 ng of an ADAR1 expression plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-PKR, anti-phospho-PKR and anti-actin antibodies.
    Quickchange Site Directed Mutagenesis Kit, supplied by Stratagene, used in various techniques. Bioz Stars score: 93/100, based on 26650 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Hitachi Ltd 3730 xl abi hitachi sequencer
    ADAR1 proviral effect is mediated through inhibition of <t>PKR</t> activation. 293-T cells were transfected with 4 μg of (A) pACH or (B) pH6neo molecular clones and increasing amount (0, 5, 50, 500 ng) of ADAR1 plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-Tax1, anti-Tax2, anti-PKR, anti-phospho-PKR and anti-actin antibodies. (C, D) : 293-T cells were transfected with <t>siRNA</t> directed against PKR (20nM) or with control siRNA. Twelve hours later cells were transfected with 20nM of the same siRNA together with 1.2 μg of (C) pACH, (D) pH6neo and 125 ng of an ADAR1 expression plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-PKR, anti-phospho-PKR and anti-actin antibodies.
    3730 Xl Abi Hitachi Sequencer, supplied by Hitachi Ltd, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Corning Life Sciences 96 well format transwell chambers
    ADAR1 proviral effect is mediated through inhibition of <t>PKR</t> activation. 293-T cells were transfected with 4 μg of (A) pACH or (B) pH6neo molecular clones and increasing amount (0, 5, 50, 500 ng) of ADAR1 plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-Tax1, anti-Tax2, anti-PKR, anti-phospho-PKR and anti-actin antibodies. (C, D) : 293-T cells were transfected with <t>siRNA</t> directed against PKR (20nM) or with control siRNA. Twelve hours later cells were transfected with 20nM of the same siRNA together with 1.2 μg of (C) pACH, (D) pH6neo and 125 ng of an ADAR1 expression plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-PKR, anti-phospho-PKR and anti-actin antibodies.
    96 Well Format Transwell Chambers, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 85/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher 96 well optical bottom plates
    ADAR1 proviral effect is mediated through inhibition of <t>PKR</t> activation. 293-T cells were transfected with 4 μg of (A) pACH or (B) pH6neo molecular clones and increasing amount (0, 5, 50, 500 ng) of ADAR1 plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-Tax1, anti-Tax2, anti-PKR, anti-phospho-PKR and anti-actin antibodies. (C, D) : 293-T cells were transfected with <t>siRNA</t> directed against PKR (20nM) or with control siRNA. Twelve hours later cells were transfected with 20nM of the same siRNA together with 1.2 μg of (C) pACH, (D) pH6neo and 125 ng of an ADAR1 expression plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-PKR, anti-phospho-PKR and anti-actin antibodies.
    96 Well Optical Bottom Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 111 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher abi prism r 3100 sequencing system
    ADAR1 proviral effect is mediated through inhibition of <t>PKR</t> activation. 293-T cells were transfected with 4 μg of (A) pACH or (B) pH6neo molecular clones and increasing amount (0, 5, 50, 500 ng) of ADAR1 plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-Tax1, anti-Tax2, anti-PKR, anti-phospho-PKR and anti-actin antibodies. (C, D) : 293-T cells were transfected with <t>siRNA</t> directed against PKR (20nM) or with control siRNA. Twelve hours later cells were transfected with 20nM of the same siRNA together with 1.2 μg of (C) pACH, (D) pH6neo and 125 ng of an ADAR1 expression plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-PKR, anti-phospho-PKR and anti-actin antibodies.
    Abi Prism R 3100 Sequencing System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    PAA Laboratories accutase
    ADAR1 proviral effect is mediated through inhibition of <t>PKR</t> activation. 293-T cells were transfected with 4 μg of (A) pACH or (B) pH6neo molecular clones and increasing amount (0, 5, 50, 500 ng) of ADAR1 plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-Tax1, anti-Tax2, anti-PKR, anti-phospho-PKR and anti-actin antibodies. (C, D) : 293-T cells were transfected with <t>siRNA</t> directed against PKR (20nM) or with control siRNA. Twelve hours later cells were transfected with 20nM of the same siRNA together with 1.2 μg of (C) pACH, (D) pH6neo and 125 ng of an ADAR1 expression plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-PKR, anti-phospho-PKR and anti-actin antibodies.
    Accutase, supplied by PAA Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 1445 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Applied Photophysics advanced solar photophysics
    ADAR1 proviral effect is mediated through inhibition of <t>PKR</t> activation. 293-T cells were transfected with 4 μg of (A) pACH or (B) pH6neo molecular clones and increasing amount (0, 5, 50, 500 ng) of ADAR1 plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-Tax1, anti-Tax2, anti-PKR, anti-phospho-PKR and anti-actin antibodies. (C, D) : 293-T cells were transfected with <t>siRNA</t> directed against PKR (20nM) or with control siRNA. Twelve hours later cells were transfected with 20nM of the same siRNA together with 1.2 μg of (C) pACH, (D) pH6neo and 125 ng of an ADAR1 expression plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-PKR, anti-phospho-PKR and anti-actin antibodies.
    Advanced Solar Photophysics, supplied by Applied Photophysics, used in various techniques. Bioz Stars score: 88/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ADAR1 proviral effect is mediated through inhibition of <t>PKR</t> activation. 293-T cells were transfected with 4 μg of (A) pACH or (B) pH6neo molecular clones and increasing amount (0, 5, 50, 500 ng) of ADAR1 plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-Tax1, anti-Tax2, anti-PKR, anti-phospho-PKR and anti-actin antibodies. (C, D) : 293-T cells were transfected with <t>siRNA</t> directed against PKR (20nM) or with control siRNA. Twelve hours later cells were transfected with 20nM of the same siRNA together with 1.2 μg of (C) pACH, (D) pH6neo and 125 ng of an ADAR1 expression plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-PKR, anti-phospho-PKR and anti-actin antibodies.
    Alexa 647 α Bgt, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ADAR1 proviral effect is mediated through inhibition of <t>PKR</t> activation. 293-T cells were transfected with 4 μg of (A) pACH or (B) pH6neo molecular clones and increasing amount (0, 5, 50, 500 ng) of ADAR1 plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-Tax1, anti-Tax2, anti-PKR, anti-phospho-PKR and anti-actin antibodies. (C, D) : 293-T cells were transfected with <t>siRNA</t> directed against PKR (20nM) or with control siRNA. Twelve hours later cells were transfected with 20nM of the same siRNA together with 1.2 μg of (C) pACH, (D) pH6neo and 125 ng of an ADAR1 expression plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-PKR, anti-phospho-PKR and anti-actin antibodies.
    Alexa Fluor 680 Conjugated Wheat Germ Agglutinin Wga, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Model of HIV-1 transcriptional activation by Tat and RelB. HIV-1 Tat interacts with RelB. RelB and Tat are recruited to LTR DNA to assist transcription, which increases recruitment of RNA Pol II to the LTR and leads to greater expression of viral genes

    Journal: Retrovirology

    Article Title: Cellular RelB interacts with the transactivator Tat and enhance HIV-1 expression

    doi: 10.1186/s12977-018-0447-9

    Figure Lengend Snippet: Model of HIV-1 transcriptional activation by Tat and RelB. HIV-1 Tat interacts with RelB. RelB and Tat are recruited to LTR DNA to assist transcription, which increases recruitment of RNA Pol II to the LTR and leads to greater expression of viral genes

    Article Snippet: Pache L, Dutra MS, Spivak AM, Marlett JM, Murry JP, Hwang Y, Maestre AM, Manganaro L, Vamos M, Teriete P, et al. BIRC2/cIAP1 is a negative regulator of HIV-1 transcription and can be targeted by Smac mimetics to promote reversal of viral latency.

    Techniques: Activation Assay, Expressing

    Requirement of RelB for the activation of latent HIV-1 in J-Lat A72 cells. a Illustration of the J-Lat A72 latency model. b Western blots to examine the overexpressed RelB or RelB knockout in J-Lat A72 cell lines. c RelB enhanced Tat-mediated activation of latent HIV-1 in J-Lat A72 cells. J-Lat A72 cells (2 × 10 6 ) were transduced with VSV-G pseudotyped lentiviral VLPs expressing Tat. Twenty-four hours post infection, cells were treated with 10 nM PMA or DMSO for 24 h, then subjected to flow cytometry to detect GFP expressing cells using a FACSCalibur flow cytometer. Data were analyzed using the FlowJo software. d Analysis of HIV-1 LTR transcriptional activation by flow cytometry. Results from three independent experiments were summarized

    Journal: Retrovirology

    Article Title: Cellular RelB interacts with the transactivator Tat and enhance HIV-1 expression

    doi: 10.1186/s12977-018-0447-9

    Figure Lengend Snippet: Requirement of RelB for the activation of latent HIV-1 in J-Lat A72 cells. a Illustration of the J-Lat A72 latency model. b Western blots to examine the overexpressed RelB or RelB knockout in J-Lat A72 cell lines. c RelB enhanced Tat-mediated activation of latent HIV-1 in J-Lat A72 cells. J-Lat A72 cells (2 × 10 6 ) were transduced with VSV-G pseudotyped lentiviral VLPs expressing Tat. Twenty-four hours post infection, cells were treated with 10 nM PMA or DMSO for 24 h, then subjected to flow cytometry to detect GFP expressing cells using a FACSCalibur flow cytometer. Data were analyzed using the FlowJo software. d Analysis of HIV-1 LTR transcriptional activation by flow cytometry. Results from three independent experiments were summarized

    Article Snippet: Pache L, Dutra MS, Spivak AM, Marlett JM, Murry JP, Hwang Y, Maestre AM, Manganaro L, Vamos M, Teriete P, et al. BIRC2/cIAP1 is a negative regulator of HIV-1 transcription and can be targeted by Smac mimetics to promote reversal of viral latency.

    Techniques: Activation Assay, Western Blot, Knock-Out, Transduction, Expressing, Infection, Flow Cytometry, Cytometry, Software

    Association of RelB and Tat binding to the HIV-1 LTR. a Schematic illustration of the HIV-1 LTR and location of oligonucleotide primers. ChIP primers for HIV-1 LTR analysis were designed to span the enhancer region, including the duplicated κB enhancers (− 104 to − 81) and specificity protein 1 binding sites (SP1, − 79 to − 47). Primers for analysis of initiated HIV-1 transcripts were directed against TAR. b – d TZM-bl cells were transfected with pFlag-Tat (6 μg) or vector plasmids. The relative expression of Tat and RelB in the transfected cells was monitored by a western blot assay ( b ). Chromatin of the fixed TZM-bl cells (1 × 10 7 ) was immunoprecipitated with the indicated antibodies. Samples were assessed for enrichment of HIV-1 LTR DNA by PCR ( c ). Relative levels of the LTR DNA that were co-precipitated with RNA Pol II, RelB, and p52 were quantified by real-time PCR performed in triplicate. The data are normalized to the input controls. Data are representative of three independent experiments ( d ). e – h Control and RELB − / − TZM-bl cells were transfected with pFlag-Tat (6 μg) or vector plasmids ( e ). The co-precipitated LTR DNA was assessed by PCR ( f ). Tat ( g ) and RNA Pol II ( h ) associated HIV-1 LTR was quantified by real-time PCR

    Journal: Retrovirology

    Article Title: Cellular RelB interacts with the transactivator Tat and enhance HIV-1 expression

    doi: 10.1186/s12977-018-0447-9

    Figure Lengend Snippet: Association of RelB and Tat binding to the HIV-1 LTR. a Schematic illustration of the HIV-1 LTR and location of oligonucleotide primers. ChIP primers for HIV-1 LTR analysis were designed to span the enhancer region, including the duplicated κB enhancers (− 104 to − 81) and specificity protein 1 binding sites (SP1, − 79 to − 47). Primers for analysis of initiated HIV-1 transcripts were directed against TAR. b – d TZM-bl cells were transfected with pFlag-Tat (6 μg) or vector plasmids. The relative expression of Tat and RelB in the transfected cells was monitored by a western blot assay ( b ). Chromatin of the fixed TZM-bl cells (1 × 10 7 ) was immunoprecipitated with the indicated antibodies. Samples were assessed for enrichment of HIV-1 LTR DNA by PCR ( c ). Relative levels of the LTR DNA that were co-precipitated with RNA Pol II, RelB, and p52 were quantified by real-time PCR performed in triplicate. The data are normalized to the input controls. Data are representative of three independent experiments ( d ). e – h Control and RELB − / − TZM-bl cells were transfected with pFlag-Tat (6 μg) or vector plasmids ( e ). The co-precipitated LTR DNA was assessed by PCR ( f ). Tat ( g ) and RNA Pol II ( h ) associated HIV-1 LTR was quantified by real-time PCR

    Article Snippet: Pache L, Dutra MS, Spivak AM, Marlett JM, Murry JP, Hwang Y, Maestre AM, Manganaro L, Vamos M, Teriete P, et al. BIRC2/cIAP1 is a negative regulator of HIV-1 transcription and can be targeted by Smac mimetics to promote reversal of viral latency.

    Techniques: Binding Assay, Chromatin Immunoprecipitation, Transfection, Plasmid Preparation, Expressing, Western Blot, Immunoprecipitation, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

    RelB increases HIV-1 gene transcription. a HEK 293T cells (0.5 × 10 6 ) in a 6-well plate were transfected with increasing amounts of pHA-RelB, together with pVSV-G (0.3 μg) and pNL4-3.Luc.env − . (1 μg). Forty-eight hours post transfection, cells were collected for western blotting and luciferase activity analysis. b Stable RelB-overexpressing cells, RELB − / − cells, and control HEK 293T cell lines (0.5 × 10 6 ) were transfected with pVSV-G (0.3 μg) and pNL4-3.Luc.env − (1 μg). Western blotting and Luciferase activity essays were performed to assess viral gene expression. c Control and RELB − / − HeLa cells were transfected with pHA-RelB (500 ng) or vector DNA. Twenty-four hours post transfection, cells were infected with VSVG-NL4-3.Luc.env − equivalent to 20 ng p24 for 40 h. Cells were collected for luciferase assay and western blotting analyses. d and e RelB overexpressing or RELB − / − Jurkat cells were infected with VSVG-NLENY1-ES-IRES equivalent to 20 ng p24 for 48 h. Gating on live Jurkat cells was based on size (FSC-H) and granularity (SSC-H). YFP positive cells were scored by flow cytometry 48 h after infection ( d ). Results from three independent experiments were summarized, and cells were collected for western blotting analysis ( e ). f PBMCs (2 × 10 6 ) were infected with RelB-expressing VLPs, or transfected with siRNA. After 24 h, infection with VSV-G pseudotyped NL4-3.Luc.env − (equivalent to 500 ng p24) was performed by spinoculation at 300× g for 30 min. After 24 h, whole cell lysates were examined in luciferase assay and western blotting with the indicated antibodies

    Journal: Retrovirology

    Article Title: Cellular RelB interacts with the transactivator Tat and enhance HIV-1 expression

    doi: 10.1186/s12977-018-0447-9

    Figure Lengend Snippet: RelB increases HIV-1 gene transcription. a HEK 293T cells (0.5 × 10 6 ) in a 6-well plate were transfected with increasing amounts of pHA-RelB, together with pVSV-G (0.3 μg) and pNL4-3.Luc.env − . (1 μg). Forty-eight hours post transfection, cells were collected for western blotting and luciferase activity analysis. b Stable RelB-overexpressing cells, RELB − / − cells, and control HEK 293T cell lines (0.5 × 10 6 ) were transfected with pVSV-G (0.3 μg) and pNL4-3.Luc.env − (1 μg). Western blotting and Luciferase activity essays were performed to assess viral gene expression. c Control and RELB − / − HeLa cells were transfected with pHA-RelB (500 ng) or vector DNA. Twenty-four hours post transfection, cells were infected with VSVG-NL4-3.Luc.env − equivalent to 20 ng p24 for 40 h. Cells were collected for luciferase assay and western blotting analyses. d and e RelB overexpressing or RELB − / − Jurkat cells were infected with VSVG-NLENY1-ES-IRES equivalent to 20 ng p24 for 48 h. Gating on live Jurkat cells was based on size (FSC-H) and granularity (SSC-H). YFP positive cells were scored by flow cytometry 48 h after infection ( d ). Results from three independent experiments were summarized, and cells were collected for western blotting analysis ( e ). f PBMCs (2 × 10 6 ) were infected with RelB-expressing VLPs, or transfected with siRNA. After 24 h, infection with VSV-G pseudotyped NL4-3.Luc.env − (equivalent to 500 ng p24) was performed by spinoculation at 300× g for 30 min. After 24 h, whole cell lysates were examined in luciferase assay and western blotting with the indicated antibodies

    Article Snippet: Pache L, Dutra MS, Spivak AM, Marlett JM, Murry JP, Hwang Y, Maestre AM, Manganaro L, Vamos M, Teriete P, et al. BIRC2/cIAP1 is a negative regulator of HIV-1 transcription and can be targeted by Smac mimetics to promote reversal of viral latency.

    Techniques: Transfection, Western Blot, Luciferase, Activity Assay, Expressing, Plasmid Preparation, Infection, Flow Cytometry, Cytometry

    The κB elements are required for RelB to enhance Tat-mediated transcription of HIV-1 LTR. a Schematic representation of 5′-truncated pGL3-HIV-1-LTR-deletion reporter constructs. TAR (+ 1 to + 60), SP1 (− 79 to − 47), NF-κB (− 104 to − 81), and NRE (− 340 to − 184) sites are indicated. b Responsiveness of 5′-truncated pGL3-HIV-1-LTR reporter constructs to Tat and RelB. HEK 293T cells (0.2 × 10 6 ) were cotransfected with each of the 5′-truncated pGL3-HIV-1-LTR-deletion reporter constructs (100 ng) in the presence of Tat-expressing plasmids, together with pCMV-RelB constructs or control vectors. c HeLa cells (0.1 × 10 6 ) were transfected with the indicated HIV-1 LTR-Luc reporter plasmids (100 ng) and Tat-expressing plasmids (100 ng), together with pCMV-RelB constructs (100 ng) or control vectors. To normalize transfection efficiency, phRluc-TK (1 ng) was co-transfected. Luciferase assays were performed 48 h post transfection. The Rel. Luc. Act in the figure was calculated relative to normalized luciferase activity of pHIV-1-LTR-luc with Tat (with or without RelB) and vectors co-transfected cells

    Journal: Retrovirology

    Article Title: Cellular RelB interacts with the transactivator Tat and enhance HIV-1 expression

    doi: 10.1186/s12977-018-0447-9

    Figure Lengend Snippet: The κB elements are required for RelB to enhance Tat-mediated transcription of HIV-1 LTR. a Schematic representation of 5′-truncated pGL3-HIV-1-LTR-deletion reporter constructs. TAR (+ 1 to + 60), SP1 (− 79 to − 47), NF-κB (− 104 to − 81), and NRE (− 340 to − 184) sites are indicated. b Responsiveness of 5′-truncated pGL3-HIV-1-LTR reporter constructs to Tat and RelB. HEK 293T cells (0.2 × 10 6 ) were cotransfected with each of the 5′-truncated pGL3-HIV-1-LTR-deletion reporter constructs (100 ng) in the presence of Tat-expressing plasmids, together with pCMV-RelB constructs or control vectors. c HeLa cells (0.1 × 10 6 ) were transfected with the indicated HIV-1 LTR-Luc reporter plasmids (100 ng) and Tat-expressing plasmids (100 ng), together with pCMV-RelB constructs (100 ng) or control vectors. To normalize transfection efficiency, phRluc-TK (1 ng) was co-transfected. Luciferase assays were performed 48 h post transfection. The Rel. Luc. Act in the figure was calculated relative to normalized luciferase activity of pHIV-1-LTR-luc with Tat (with or without RelB) and vectors co-transfected cells

    Article Snippet: Pache L, Dutra MS, Spivak AM, Marlett JM, Murry JP, Hwang Y, Maestre AM, Manganaro L, Vamos M, Teriete P, et al. BIRC2/cIAP1 is a negative regulator of HIV-1 transcription and can be targeted by Smac mimetics to promote reversal of viral latency.

    Techniques: Construct, Expressing, Transfection, Luciferase, Activated Clotting Time Assay, Activity Assay

    NF-κB elements are essential for RelB to enhance Tat recruitment to the LTR. a–d HeLa cells (1 × 10 7 ) were transfected with pHIV-1-LTR-luc (6 μg) or pHIV-1-LTR mutNF-κB (6 μg), together with pFlag-Tat (6 μg) or vector plasmids. The relative expression of Tat and RelB in the transfected cells was monitored by western blots ( a ). Chromatin of the fixed cells was immunoprecipitated with the indicated antibodies. Samples were assessed for enrichment of HIV-1 LTR DNA by PCR ( b ). Tat ( c ) and RNA Pol II ( d ) associated HIV-1 LTR DNA was quantified by real-time PCR. (E to H) Control and RELB − / − HeLa cells (1 × 10 7 ) were transfected with pHIV-1-LTR mutNF-κB (6 μg), together with pFlag-Tat (6 μg) or vector plasmids ( e ). Samples were assessed for enrichment of HIV-1 LTR DNA by PCR. f Tat ( g ) and RNA Pol II ( h ) enriched HIV-1 LTR DNA was quantified by real-time PCR

    Journal: Retrovirology

    Article Title: Cellular RelB interacts with the transactivator Tat and enhance HIV-1 expression

    doi: 10.1186/s12977-018-0447-9

    Figure Lengend Snippet: NF-κB elements are essential for RelB to enhance Tat recruitment to the LTR. a–d HeLa cells (1 × 10 7 ) were transfected with pHIV-1-LTR-luc (6 μg) or pHIV-1-LTR mutNF-κB (6 μg), together with pFlag-Tat (6 μg) or vector plasmids. The relative expression of Tat and RelB in the transfected cells was monitored by western blots ( a ). Chromatin of the fixed cells was immunoprecipitated with the indicated antibodies. Samples were assessed for enrichment of HIV-1 LTR DNA by PCR ( b ). Tat ( c ) and RNA Pol II ( d ) associated HIV-1 LTR DNA was quantified by real-time PCR. (E to H) Control and RELB − / − HeLa cells (1 × 10 7 ) were transfected with pHIV-1-LTR mutNF-κB (6 μg), together with pFlag-Tat (6 μg) or vector plasmids ( e ). Samples were assessed for enrichment of HIV-1 LTR DNA by PCR. f Tat ( g ) and RNA Pol II ( h ) enriched HIV-1 LTR DNA was quantified by real-time PCR

    Article Snippet: Pache L, Dutra MS, Spivak AM, Marlett JM, Murry JP, Hwang Y, Maestre AM, Manganaro L, Vamos M, Teriete P, et al. BIRC2/cIAP1 is a negative regulator of HIV-1 transcription and can be targeted by Smac mimetics to promote reversal of viral latency.

    Techniques: Transfection, Plasmid Preparation, Expressing, Western Blot, Immunoprecipitation, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

    Interaction between HIV-1 Tat and RelB. a , b Myc-Tat (3 μg) was transfected into HEK 293T cells (4 × 10 6 ) together with empty vectors (control) (3 μg) or pFlag-RelB (3 μg) Co-immunoprecipitation was performed with anti-Flag ( a ) or anti-Myc ( b ) antibodies. Samples of both cell lysates and immunoprecipitates were subjected to western blotting and probed with rabbit anti-Myc and anti-Flag antibodies. c Co-IP of endogenous RelB and ectopically expressed Tat. The lysate from HA-Tat-expressing HeLa cells (4 × 10 6 ) was immunoprecipitated with mouse anti-HA antibodies, and the precipitated proteins were examined with western blotting. d Effect of RNases on the association of endogenous RelB and ectopically expressed Tat. Lysates (in the presence or absence of RNase A [5 μg/ml]) of HA-Tat expressing HeLa cells (4 × 10 6 ) were immunoprecipitated with control rabbit IgG or rabbit anti-RelB antibodies. Samples from cell lysates and immunoprecipitates were subjected to western blotting. e Tat partially co-localizes with RelB. HeLa cells (0.1 × 10 6 ) were transfected with HA-Tat (200 ng) and Flag-RelB (200 ng) plasmid DNA. Indirect IFA was performed to detect HA-Tat (with rabbit anti-HA antibody and TRITC-conjugated goat anti rabbit secondary antibody) and Flag-RelB (with mouse anti-Flag antibody and FITC-conjugated goat anti mouse secondary antibody). Nuclei were visualized with DAPI staining. Representative images are shown. The inset shows a higher magnification of the boxed area

    Journal: Retrovirology

    Article Title: Cellular RelB interacts with the transactivator Tat and enhance HIV-1 expression

    doi: 10.1186/s12977-018-0447-9

    Figure Lengend Snippet: Interaction between HIV-1 Tat and RelB. a , b Myc-Tat (3 μg) was transfected into HEK 293T cells (4 × 10 6 ) together with empty vectors (control) (3 μg) or pFlag-RelB (3 μg) Co-immunoprecipitation was performed with anti-Flag ( a ) or anti-Myc ( b ) antibodies. Samples of both cell lysates and immunoprecipitates were subjected to western blotting and probed with rabbit anti-Myc and anti-Flag antibodies. c Co-IP of endogenous RelB and ectopically expressed Tat. The lysate from HA-Tat-expressing HeLa cells (4 × 10 6 ) was immunoprecipitated with mouse anti-HA antibodies, and the precipitated proteins were examined with western blotting. d Effect of RNases on the association of endogenous RelB and ectopically expressed Tat. Lysates (in the presence or absence of RNase A [5 μg/ml]) of HA-Tat expressing HeLa cells (4 × 10 6 ) were immunoprecipitated with control rabbit IgG or rabbit anti-RelB antibodies. Samples from cell lysates and immunoprecipitates were subjected to western blotting. e Tat partially co-localizes with RelB. HeLa cells (0.1 × 10 6 ) were transfected with HA-Tat (200 ng) and Flag-RelB (200 ng) plasmid DNA. Indirect IFA was performed to detect HA-Tat (with rabbit anti-HA antibody and TRITC-conjugated goat anti rabbit secondary antibody) and Flag-RelB (with mouse anti-Flag antibody and FITC-conjugated goat anti mouse secondary antibody). Nuclei were visualized with DAPI staining. Representative images are shown. The inset shows a higher magnification of the boxed area

    Article Snippet: Pache L, Dutra MS, Spivak AM, Marlett JM, Murry JP, Hwang Y, Maestre AM, Manganaro L, Vamos M, Teriete P, et al. BIRC2/cIAP1 is a negative regulator of HIV-1 transcription and can be targeted by Smac mimetics to promote reversal of viral latency.

    Techniques: Transfection, Immunoprecipitation, Western Blot, Co-Immunoprecipitation Assay, Expressing, Plasmid Preparation, Immunofluorescence, Staining

    Endogenous RelB facilitates Tat-induced, TAR-independent HIV-1 LTR transactivation. a Control and RELB − / − HeLa cells (0.1 × 10 6 ) were transfected by indicated plasmids (pHA-Tat: 100 ng, pHA-RelB: 100 ng) and pHIV-1-LTR-luc luciferase reporter plasmids (100 ng). b Schematic representation of the CycT1-Tat-TAR ternary complex and TAR mutation. c Control and RELB − / − HeLa cells (0.1 × 10 6 ) were transfected with the indicated plasmids (pHA-Tat: 100 ng, pHA-RelB: 100 ng) and pHIV-1-LTR-luc luciferase reporter plasmids (100 ng). To normalize transfection efficiency, phRluc-TK (1 ng) was co-transfected. The Rel. Luc. Act in the figure was calculated relative to normalized luciferase activity of pHIV-1-LTR-luc with Tat (with or without RelB) and vectors co-transfected cells. The relative expression of RelB in the transfected cells was monitored by western blotting

    Journal: Retrovirology

    Article Title: Cellular RelB interacts with the transactivator Tat and enhance HIV-1 expression

    doi: 10.1186/s12977-018-0447-9

    Figure Lengend Snippet: Endogenous RelB facilitates Tat-induced, TAR-independent HIV-1 LTR transactivation. a Control and RELB − / − HeLa cells (0.1 × 10 6 ) were transfected by indicated plasmids (pHA-Tat: 100 ng, pHA-RelB: 100 ng) and pHIV-1-LTR-luc luciferase reporter plasmids (100 ng). b Schematic representation of the CycT1-Tat-TAR ternary complex and TAR mutation. c Control and RELB − / − HeLa cells (0.1 × 10 6 ) were transfected with the indicated plasmids (pHA-Tat: 100 ng, pHA-RelB: 100 ng) and pHIV-1-LTR-luc luciferase reporter plasmids (100 ng). To normalize transfection efficiency, phRluc-TK (1 ng) was co-transfected. The Rel. Luc. Act in the figure was calculated relative to normalized luciferase activity of pHIV-1-LTR-luc with Tat (with or without RelB) and vectors co-transfected cells. The relative expression of RelB in the transfected cells was monitored by western blotting

    Article Snippet: Pache L, Dutra MS, Spivak AM, Marlett JM, Murry JP, Hwang Y, Maestre AM, Manganaro L, Vamos M, Teriete P, et al. BIRC2/cIAP1 is a negative regulator of HIV-1 transcription and can be targeted by Smac mimetics to promote reversal of viral latency.

    Techniques: Transfection, Luciferase, Mutagenesis, Activated Clotting Time Assay, Activity Assay, Expressing, Western Blot

    RelB enhances Tat transactivation of HIV-1 LTR. a Illustration of the luciferase reporter system. b – d HeLa cells (0.5 × 10 5 ) were cotransfected with a reporter construct containing the HIV-1 LTR upstream of the luciferase gene (100 ng) and indicated plasmids (100 ng). To normalize transfection efficiency, phRluc-TK (1 ng) was co-transfected. Total DNA amounts were adjusted to 500 ng with an empty vector. After 48 h, luciferase activities were measured. The Rel. Luc. Act in the figure was calculated relative to normalized luciferase activity of pHIV-1-LTR-luc with Tat (with or without RelB/p100) and vectors co-transfected cells. The histogram shows mean ± SD of at least three independent experiments. * P

    Journal: Retrovirology

    Article Title: Cellular RelB interacts with the transactivator Tat and enhance HIV-1 expression

    doi: 10.1186/s12977-018-0447-9

    Figure Lengend Snippet: RelB enhances Tat transactivation of HIV-1 LTR. a Illustration of the luciferase reporter system. b – d HeLa cells (0.5 × 10 5 ) were cotransfected with a reporter construct containing the HIV-1 LTR upstream of the luciferase gene (100 ng) and indicated plasmids (100 ng). To normalize transfection efficiency, phRluc-TK (1 ng) was co-transfected. Total DNA amounts were adjusted to 500 ng with an empty vector. After 48 h, luciferase activities were measured. The Rel. Luc. Act in the figure was calculated relative to normalized luciferase activity of pHIV-1-LTR-luc with Tat (with or without RelB/p100) and vectors co-transfected cells. The histogram shows mean ± SD of at least three independent experiments. * P

    Article Snippet: Pache L, Dutra MS, Spivak AM, Marlett JM, Murry JP, Hwang Y, Maestre AM, Manganaro L, Vamos M, Teriete P, et al. BIRC2/cIAP1 is a negative regulator of HIV-1 transcription and can be targeted by Smac mimetics to promote reversal of viral latency.

    Techniques: Luciferase, Construct, Transfection, Plasmid Preparation, Activated Clotting Time Assay, Activity Assay

    Inhibitory effect of IFN on Gag-raft association. IFN increases the detergent solubility of Gag (A). ACH-transfected 293T cells in the absence (upper panel) or presence (lower panel) of IFN were treated with or without 0.5% Triton X-100 (Tx100) on ice for 20 min before fixation. Gag membrane localization was detected by confocal analysis by using anti-p19 Gag . In the presence of Triton X-100, the punctate fluorescence pattern was largely preserved in DRMs without IFN. However, it was largely lost in DRMs from cells treated with 100 U of IFN/ml. IFN blocks Gag-raft colocalization (B). ACH-expressing cells were stained with anti-p19 Gag MAb (green) and anti-GM1 (red). p19 Gag immunofluorescence colocalized with raft-associated marker GM1 in the absence of IFN (the upper panel), whereas only a small portion of p19Gag was colocalized with GM1 following IFN treatment (the lower panel).

    Journal: Journal of Virology

    Article Title: Alpha Interferon Inhibits Human T-Cell Leukemia Virus Type 1 Assembly by Preventing Gag Interaction with Rafts

    doi: 10.1128/JVI.77.24.13389-13395.2003

    Figure Lengend Snippet: Inhibitory effect of IFN on Gag-raft association. IFN increases the detergent solubility of Gag (A). ACH-transfected 293T cells in the absence (upper panel) or presence (lower panel) of IFN were treated with or without 0.5% Triton X-100 (Tx100) on ice for 20 min before fixation. Gag membrane localization was detected by confocal analysis by using anti-p19 Gag . In the presence of Triton X-100, the punctate fluorescence pattern was largely preserved in DRMs without IFN. However, it was largely lost in DRMs from cells treated with 100 U of IFN/ml. IFN blocks Gag-raft colocalization (B). ACH-expressing cells were stained with anti-p19 Gag MAb (green) and anti-GM1 (red). p19 Gag immunofluorescence colocalized with raft-associated marker GM1 in the absence of IFN (the upper panel), whereas only a small portion of p19Gag was colocalized with GM1 following IFN treatment (the lower panel).

    Article Snippet: Immunoblot analyses were performed with monoclonal antibody (MAb) against viral p19Gag (ZeptoMetrix Corporation, Buffalo, N.Y.), and polyclonal antibody against cellular actin (Santa Cruz Biotechnology, Inc., Santa Cruz, Calif.).

    Techniques: Solubility, Transfection, Fluorescence, Expressing, Staining, Immunofluorescence, Marker

    SMAC mimetics induce the degradation of BIRC2 and XIAP in HIV-T CM . ( A ) Left , representative western blots of FAS, membrane-bound FASLG (mFASLG) and soluble FASLG (sFASLG) in T CM and HIV-T CM . Right , densitometric analysis of blots. n = 4. ( B ) Left , representative western blots of FAS, membrane-bound FASLG (mFASLG) and soluble FASLG (sFASLG) in A3.01 and ACH-2. Experiment was performed in triplicate. Right , densitometric analysis of blots. ( C ) Left , representative western blots of XIAP and BIRC2 in A3.01 and ACH-2. Experiment was performed in triplicate. Right , densitometric analysis of blots. ( D ) Left , representative western blots of XIAP and BIRC2 in T CM and HIV-T CM using antibody to XIAP, BIRC2 and ACTB. Right , densitometric analysis of blots. n = 4. ( E ) Left , representative western blots of LC3B isoforms, BECN1 and SQSTM1 in T CM and HIV-T CM . Right , densitometric analysis of blots. n = 4. ( F ) HIV-T CM were treated for 4 h with 100 nM bafilomycin A 1 . Left , representative western blots of LC3B isoforms and SQSTM1. Right , densitometric analysis of blots. n = 4. ( G ) T CM and HIV-T CM were treated for 24 h with increasing concentrations of birinapant, GDC-0152, or embelin. Left , representative western blots of XIAP and BIRC2. Right , densitometric analysis of blots. n = 4.

    Journal: Cell host & microbe

    Article Title: SMAC mimetics induce autophagy-dependent apoptosis of HIV-1-infected resting memory CD4+ T cells

    doi: 10.1016/j.chom.2018.09.007

    Figure Lengend Snippet: SMAC mimetics induce the degradation of BIRC2 and XIAP in HIV-T CM . ( A ) Left , representative western blots of FAS, membrane-bound FASLG (mFASLG) and soluble FASLG (sFASLG) in T CM and HIV-T CM . Right , densitometric analysis of blots. n = 4. ( B ) Left , representative western blots of FAS, membrane-bound FASLG (mFASLG) and soluble FASLG (sFASLG) in A3.01 and ACH-2. Experiment was performed in triplicate. Right , densitometric analysis of blots. ( C ) Left , representative western blots of XIAP and BIRC2 in A3.01 and ACH-2. Experiment was performed in triplicate. Right , densitometric analysis of blots. ( D ) Left , representative western blots of XIAP and BIRC2 in T CM and HIV-T CM using antibody to XIAP, BIRC2 and ACTB. Right , densitometric analysis of blots. n = 4. ( E ) Left , representative western blots of LC3B isoforms, BECN1 and SQSTM1 in T CM and HIV-T CM . Right , densitometric analysis of blots. n = 4. ( F ) HIV-T CM were treated for 4 h with 100 nM bafilomycin A 1 . Left , representative western blots of LC3B isoforms and SQSTM1. Right , densitometric analysis of blots. n = 4. ( G ) T CM and HIV-T CM were treated for 24 h with increasing concentrations of birinapant, GDC-0152, or embelin. Left , representative western blots of XIAP and BIRC2. Right , densitometric analysis of blots. n = 4.

    Article Snippet: BIRC2/cIAP1 Is a Negative Regulator of HIV-1 Transcription and Can Be Targeted by Smac Mimetics to Promote Reversal of Viral Latency .

    Techniques: Western Blot

    SMAC mimetics promote cell death via apoptosis. ( A ) HIV-T CM were pretreated with necrostatin-1 (Nec), z-VAD-fmk (z-VAD), wortmannin (Wort), or vehicle control for 2 h. Cell death was measured 24 h after subsequent 100 nM birinapant, 100 nM GDC-0152 or 10 μ M embelin treatment using a cell death ELISA. n = 4. ( B ) HIV-T CM were exposed to 100 nM birinapant, 100 nM GDC-0152 or 10 μ M embelin. Every 3 h for 24 h, cells were harvested, stained with ANXA5 and PI, and the frequency of ANXA5-positive or ANXA5/PI-positive labeling was recorded by flow cytometry. Data are the means of a representative donor. ( C ) T CM and HIV-T CM were treated with 100 nM birinapant, 100 nM GDC-0152, 10 μ M embelin, or vehicle for 24 h. Left , representative western blots of ATG7, ATG12–ATG5, FADD, MLKL, RIPK1, cleaved RIPK1 (cRIPK1), RIPK3, cleaved RIPK3 (cRIPK3), and SQSTM1 in T CM and HIV-T CM . Right , densitometric analysis of blots n = 4.

    Journal: Cell host & microbe

    Article Title: SMAC mimetics induce autophagy-dependent apoptosis of HIV-1-infected resting memory CD4+ T cells

    doi: 10.1016/j.chom.2018.09.007

    Figure Lengend Snippet: SMAC mimetics promote cell death via apoptosis. ( A ) HIV-T CM were pretreated with necrostatin-1 (Nec), z-VAD-fmk (z-VAD), wortmannin (Wort), or vehicle control for 2 h. Cell death was measured 24 h after subsequent 100 nM birinapant, 100 nM GDC-0152 or 10 μ M embelin treatment using a cell death ELISA. n = 4. ( B ) HIV-T CM were exposed to 100 nM birinapant, 100 nM GDC-0152 or 10 μ M embelin. Every 3 h for 24 h, cells were harvested, stained with ANXA5 and PI, and the frequency of ANXA5-positive or ANXA5/PI-positive labeling was recorded by flow cytometry. Data are the means of a representative donor. ( C ) T CM and HIV-T CM were treated with 100 nM birinapant, 100 nM GDC-0152, 10 μ M embelin, or vehicle for 24 h. Left , representative western blots of ATG7, ATG12–ATG5, FADD, MLKL, RIPK1, cleaved RIPK1 (cRIPK1), RIPK3, cleaved RIPK3 (cRIPK3), and SQSTM1 in T CM and HIV-T CM . Right , densitometric analysis of blots n = 4.

    Article Snippet: BIRC2/cIAP1 Is a Negative Regulator of HIV-1 Transcription and Can Be Targeted by Smac Mimetics to Promote Reversal of Viral Latency .

    Techniques: Enzyme-linked Immunosorbent Assay, Staining, Labeling, Flow Cytometry, Cytometry, Western Blot

    SMAC mimetics promote cell death via the formation of a CASP8-activating platform. ( A ) HIV-T CM simultaneous transduced with ATG2A and ATG2B shRNA (sh ATG2A/B ), ATG5 shRNA (sh ATG5) , ATG7 (sh ATG7 ), RUBCN (sh RUBCN ), SQSTM1 (sh SQSTM1 ), ULK1 (sh ULK1 ), or scrambled shRNA (shNS) were incubated with 100 nM birinapant, 100 nM GDC-0152, 10 μ M embelin, or vehicle. Cell death was measured after 24 h using a cell death ELISA. n . ( B ) HIV-T CM were pretreated with vehicle, bafilomycin A 1 ( Baf A 1 ), or chloroquine ( CQ ) before incubation with 100 nM birinapant, 100 nM GDC-0152 or 10 μ M embelin for 24 h. Cell death was measured using a cell death ELISA. n . ( C ) T CM and HIV-T CM were treated with 100 nM birinapant, 100 nM GDC-0152 or 10 μ M embelin for 24 h. Cells were lysed and CASP8 was immunoprecipitated (IP). The presence of ATG7, ATG12–ATG5, FADD, MLKL, cleaved RIPK1 (cRIPK1), RIPK3, cleaved RIPK3 (cRIPK3), and SQSTM1 and CASP8 was assayed by western blot. Left , representative western blots. Right , densitometric analysis of blots. n = 4.

    Journal: Cell host & microbe

    Article Title: SMAC mimetics induce autophagy-dependent apoptosis of HIV-1-infected resting memory CD4+ T cells

    doi: 10.1016/j.chom.2018.09.007

    Figure Lengend Snippet: SMAC mimetics promote cell death via the formation of a CASP8-activating platform. ( A ) HIV-T CM simultaneous transduced with ATG2A and ATG2B shRNA (sh ATG2A/B ), ATG5 shRNA (sh ATG5) , ATG7 (sh ATG7 ), RUBCN (sh RUBCN ), SQSTM1 (sh SQSTM1 ), ULK1 (sh ULK1 ), or scrambled shRNA (shNS) were incubated with 100 nM birinapant, 100 nM GDC-0152, 10 μ M embelin, or vehicle. Cell death was measured after 24 h using a cell death ELISA. n . ( B ) HIV-T CM were pretreated with vehicle, bafilomycin A 1 ( Baf A 1 ), or chloroquine ( CQ ) before incubation with 100 nM birinapant, 100 nM GDC-0152 or 10 μ M embelin for 24 h. Cell death was measured using a cell death ELISA. n . ( C ) T CM and HIV-T CM were treated with 100 nM birinapant, 100 nM GDC-0152 or 10 μ M embelin for 24 h. Cells were lysed and CASP8 was immunoprecipitated (IP). The presence of ATG7, ATG12–ATG5, FADD, MLKL, cleaved RIPK1 (cRIPK1), RIPK3, cleaved RIPK3 (cRIPK3), and SQSTM1 and CASP8 was assayed by western blot. Left , representative western blots. Right , densitometric analysis of blots. n = 4.

    Article Snippet: BIRC2/cIAP1 Is a Negative Regulator of HIV-1 Transcription and Can Be Targeted by Smac Mimetics to Promote Reversal of Viral Latency .

    Techniques: Transduction, shRNA, Incubation, Enzyme-linked Immunosorbent Assay, Immunoprecipitation, Western Blot

    SMAC mimetics preferentially induce cell death in HIV-T CM . ( A ) T CM and HIV-T CM were treated with SM or 1 μ . DNA was extracted from HIV-T CM . n = 4. ( B ) ELISA performed for HIV p24 antigen in supernatants from cells treated in A . n = 4. ( C ) RT-qPCR performed for extracellular release of HIV gag mRNA from cells treated in A . n = 4. ( D ) T CM and HIV-T CM were treated with SM for 24 h. Top , representative western blots of PARP1 and CASP8 cleavage (cPARP1 and cCASP8) using antibody to PARP1, CASP8, and ACTB. Bottom , densitometric analysis of blots, n = 4. ( E ) T CM and HIV-T CM . n = 4. ( F ) HIV-T CM were pretreated with vehicle control or TNF neutralizing antibody 2 h before incubation with SM for 24 h. Cell death was measured using a cell death ELISA. n = 4. ( G ) Resting CD4+ T cells were isolated from HIV infected donors on suppressive antiretroviral therapy, viral load

    Journal: Cell host & microbe

    Article Title: SMAC mimetics induce autophagy-dependent apoptosis of HIV-1-infected resting memory CD4+ T cells

    doi: 10.1016/j.chom.2018.09.007

    Figure Lengend Snippet: SMAC mimetics preferentially induce cell death in HIV-T CM . ( A ) T CM and HIV-T CM were treated with SM or 1 μ . DNA was extracted from HIV-T CM . n = 4. ( B ) ELISA performed for HIV p24 antigen in supernatants from cells treated in A . n = 4. ( C ) RT-qPCR performed for extracellular release of HIV gag mRNA from cells treated in A . n = 4. ( D ) T CM and HIV-T CM were treated with SM for 24 h. Top , representative western blots of PARP1 and CASP8 cleavage (cPARP1 and cCASP8) using antibody to PARP1, CASP8, and ACTB. Bottom , densitometric analysis of blots, n = 4. ( E ) T CM and HIV-T CM . n = 4. ( F ) HIV-T CM were pretreated with vehicle control or TNF neutralizing antibody 2 h before incubation with SM for 24 h. Cell death was measured using a cell death ELISA. n = 4. ( G ) Resting CD4+ T cells were isolated from HIV infected donors on suppressive antiretroviral therapy, viral load

    Article Snippet: BIRC2/cIAP1 Is a Negative Regulator of HIV-1 Transcription and Can Be Targeted by Smac Mimetics to Promote Reversal of Viral Latency .

    Techniques: Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot, Incubation, Isolation, Infection

    SMAC mimetics induce autophagy in HIV-T CM . ( A ) T CM and HIV-T CM were treated for 24 h with SM. Left , representative western blots of LC3B isoforms, BECN1 and SQSTM1. Right , densitometric analysis of blots. n = 4. ( B ) HIV-T CM were pretreated with bafilomycin A 1 before incubation with SM for 24 h. Left , representative western blots. Right , densitometric analysis of blots. n = 4.

    Journal: Cell host & microbe

    Article Title: SMAC mimetics induce autophagy-dependent apoptosis of HIV-1-infected resting memory CD4+ T cells

    doi: 10.1016/j.chom.2018.09.007

    Figure Lengend Snippet: SMAC mimetics induce autophagy in HIV-T CM . ( A ) T CM and HIV-T CM were treated for 24 h with SM. Left , representative western blots of LC3B isoforms, BECN1 and SQSTM1. Right , densitometric analysis of blots. n = 4. ( B ) HIV-T CM were pretreated with bafilomycin A 1 before incubation with SM for 24 h. Left , representative western blots. Right , densitometric analysis of blots. n = 4.

    Article Snippet: BIRC2/cIAP1 Is a Negative Regulator of HIV-1 Transcription and Can Be Targeted by Smac Mimetics to Promote Reversal of Viral Latency .

    Techniques: Western Blot, Incubation

    SMAC mimetics promote cell death via the autophagy dependent formation of a CASP8-activating platform involving SQSTM1. ( A ) HIV-T CM transduced with ATG5 shRNA (sh ATG5) , ATG7 (sh ATG7 . n = 4. ( B . n = 4. ( C ) HIV-T CM transduced with SQSTM1 shRNA (sh SQSTM1) . n = 4. ( D . n = 4.

    Journal: Cell host & microbe

    Article Title: SMAC mimetics induce autophagy-dependent apoptosis of HIV-1-infected resting memory CD4+ T cells

    doi: 10.1016/j.chom.2018.09.007

    Figure Lengend Snippet: SMAC mimetics promote cell death via the autophagy dependent formation of a CASP8-activating platform involving SQSTM1. ( A ) HIV-T CM transduced with ATG5 shRNA (sh ATG5) , ATG7 (sh ATG7 . n = 4. ( B . n = 4. ( C ) HIV-T CM transduced with SQSTM1 shRNA (sh SQSTM1) . n = 4. ( D . n = 4.

    Article Snippet: BIRC2/cIAP1 Is a Negative Regulator of HIV-1 Transcription and Can Be Targeted by Smac Mimetics to Promote Reversal of Viral Latency .

    Techniques: Transduction, shRNA

    ADAR1 proviral effect is mediated through inhibition of PKR activation. 293-T cells were transfected with 4 μg of (A) pACH or (B) pH6neo molecular clones and increasing amount (0, 5, 50, 500 ng) of ADAR1 plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-Tax1, anti-Tax2, anti-PKR, anti-phospho-PKR and anti-actin antibodies. (C, D) : 293-T cells were transfected with siRNA directed against PKR (20nM) or with control siRNA. Twelve hours later cells were transfected with 20nM of the same siRNA together with 1.2 μg of (C) pACH, (D) pH6neo and 125 ng of an ADAR1 expression plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-PKR, anti-phospho-PKR and anti-actin antibodies.

    Journal: Retrovirology

    Article Title: ADAR1 enhances HTLV-1 and HTLV-2 replication through inhibition of PKR activity

    doi: 10.1186/s12977-014-0093-9

    Figure Lengend Snippet: ADAR1 proviral effect is mediated through inhibition of PKR activation. 293-T cells were transfected with 4 μg of (A) pACH or (B) pH6neo molecular clones and increasing amount (0, 5, 50, 500 ng) of ADAR1 plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-Tax1, anti-Tax2, anti-PKR, anti-phospho-PKR and anti-actin antibodies. (C, D) : 293-T cells were transfected with siRNA directed against PKR (20nM) or with control siRNA. Twelve hours later cells were transfected with 20nM of the same siRNA together with 1.2 μg of (C) pACH, (D) pH6neo and 125 ng of an ADAR1 expression plasmid. Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24 gag , anti-PKR, anti-phospho-PKR and anti-actin antibodies.

    Article Snippet: The following day, 20 nM of PKR siRNA (ON-TARGETplus SMART pool EIF2AK2, Fermentas) or control siRNA (ON-TARGETplus Non-targeting Pool, Fermentas) were transfected (HiPerfect reagent, Qiagen) as previously described [ ].

    Techniques: Inhibition, Activation Assay, Transfection, Clone Assay, Plasmid Preparation, Western Blot, Expressing