lgtv replicons Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    New England Biolabs ecorv restriction endonuclease
    Ecorv Restriction Endonuclease, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ecorv restriction endonuclease/product/New England Biolabs
    Average 99 stars, based on 22 article reviews
    Price from $9.99 to $1999.99
    ecorv restriction endonuclease - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher mmessage mmachine kit
    Mmessage Mmachine Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 11610 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mmessage mmachine kit/product/Thermo Fisher
    Average 99 stars, based on 11610 article reviews
    Price from $9.99 to $1999.99
    mmessage mmachine kit - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    95
    Millipore sv40 large t antigen
    Sv40 Large T Antigen, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 260 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sv40 large t antigen/product/Millipore
    Average 95 stars, based on 260 article reviews
    Price from $9.99 to $1999.99
    sv40 large t antigen - by Bioz Stars, 2020-08
    95/100 stars
      Buy from Supplier

    99
    Millipore spectinomycin
    Spectinomycin, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1295 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/spectinomycin/product/Millipore
    Average 99 stars, based on 1295 article reviews
    Price from $9.99 to $1999.99
    spectinomycin - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    98
    Taconic Biosciences scid mice
    Scid Mice, supplied by Taconic Biosciences, used in various techniques. Bioz Stars score: 98/100, based on 460 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/scid mice/product/Taconic Biosciences
    Average 98 stars, based on 460 article reviews
    Price from $9.99 to $1999.99
    scid mice - by Bioz Stars, 2020-08
    98/100 stars
      Buy from Supplier

    99
    Thermo Fisher superscript iii first strand kit
    Superscript Iii First Strand Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2815 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/superscript iii first strand kit/product/Thermo Fisher
    Average 99 stars, based on 2815 article reviews
    Price from $9.99 to $1999.99
    superscript iii first strand kit - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    88
    Agilent technologies ide8 cells
    Analysis of Ago and Dcr protein-encoding genes in the Ixodes scapularis genome. ( A ) and ( B ) are gene trees for metazoan Ago-subfamily genes (A) and Dcr genes (B) respectively, constructed using a Bayesian approach under a GTR model (nodes are labeled if they receive > 90% support; see Materials and Methods). Trees are unrooted, but presented as if the root fell between the two Cnidarian homologs. ( C ) Detection of transcripts encoding Ago and Dcr proteins in <t>IDE8</t> cells by RT-PCR. RNA not treated with reverse transcriptase was used as control during the PCR reaction (no RT). See also Supplementary Figure S5.
    Ide8 Cells, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 88/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ide8 cells/product/Agilent technologies
    Average 88 stars, based on 27 article reviews
    Price from $9.99 to $1999.99
    ide8 cells - by Bioz Stars, 2020-08
    88/100 stars
      Buy from Supplier

    99
    Bio-Rad iq sybr green supermix
    Analysis of Ago and Dcr protein-encoding genes in the Ixodes scapularis genome. ( A ) and ( B ) are gene trees for metazoan Ago-subfamily genes (A) and Dcr genes (B) respectively, constructed using a Bayesian approach under a GTR model (nodes are labeled if they receive > 90% support; see Materials and Methods). Trees are unrooted, but presented as if the root fell between the two Cnidarian homologs. ( C ) Detection of transcripts encoding Ago and Dcr proteins in <t>IDE8</t> cells by RT-PCR. RNA not treated with reverse transcriptase was used as control during the PCR reaction (no RT). See also Supplementary Figure S5.
    Iq Sybr Green Supermix, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 72789 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/iq sybr green supermix/product/Bio-Rad
    Average 99 stars, based on 72789 article reviews
    Price from $9.99 to $1999.99
    iq sybr green supermix - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    92
    Exosome Diagnostics unknown pfu
    Analysis of Ago and Dcr protein-encoding genes in the Ixodes scapularis genome. ( A ) and ( B ) are gene trees for metazoan Ago-subfamily genes (A) and Dcr genes (B) respectively, constructed using a Bayesian approach under a GTR model (nodes are labeled if they receive > 90% support; see Materials and Methods). Trees are unrooted, but presented as if the root fell between the two Cnidarian homologs. ( C ) Detection of transcripts encoding Ago and Dcr proteins in <t>IDE8</t> cells by RT-PCR. RNA not treated with reverse transcriptase was used as control during the PCR reaction (no RT). See also Supplementary Figure S5.
    Unknown Pfu, supplied by Exosome Diagnostics, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/unknown pfu/product/Exosome Diagnostics
    Average 92 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    unknown pfu - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    92
    Exosome Diagnostics pbs
    Analysis of Ago and Dcr protein-encoding genes in the Ixodes scapularis genome. ( A ) and ( B ) are gene trees for metazoan Ago-subfamily genes (A) and Dcr genes (B) respectively, constructed using a Bayesian approach under a GTR model (nodes are labeled if they receive > 90% support; see Materials and Methods). Trees are unrooted, but presented as if the root fell between the two Cnidarian homologs. ( C ) Detection of transcripts encoding Ago and Dcr proteins in <t>IDE8</t> cells by RT-PCR. RNA not treated with reverse transcriptase was used as control during the PCR reaction (no RT). See also Supplementary Figure S5.
    Pbs, supplied by Exosome Diagnostics, used in various techniques. Bioz Stars score: 92/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbs/product/Exosome Diagnostics
    Average 92 stars, based on 44 article reviews
    Price from $9.99 to $1999.99
    pbs - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    92
    Exosome Diagnostics n2a cells
    Endothelial cell-derived exosomes mediate transmission of LGTV RNA from bEnd.3 cells to <t>N2a</t> neuronal cells. The bEnd.3 cells (1 x 10 5 ) were infected with 6 MOI of LGTV. QRT-PCR analysis showing total LGTV loads (A) or levels of LGTV positive-sense strand or negative-sense strand (B) in exosomes isolated from bEnd.3 cells at different time points (24, 48, 72, 96 and 120 h p.i.). (C) Viral loads were determined after 24 h p.i. by the presence of LGTV in N2a cells (1 x 10 5 ) treated with LGTV containing exosomes (20 μl) or supernatant (400 μl) fractions prepared from 24 or 48 h p.i. of bEnd.3 cells. (D) Viral loads at 48 h p.i. of N2a cells (plated in lower chamber) as determined by the presence of LGTV in a transwell assay performed with 1x 10 5 of bEnd.3 cells treated with 20 μl brain endothelial cell-derived exosomes (in upper chamber) for 4 h in the presence or absence of exosome inhibitor (5 μM) (in upper chamber) or infected with LGTV laboratory virus stocks is shown. LGTV transcript levels were normalized to mouse beta-actin. P value determined by Student’s two-tail t test is shown. Representative data is shown from three independent experiments.
    N2a Cells, supplied by Exosome Diagnostics, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/n2a cells/product/Exosome Diagnostics
    Average 92 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    n2a cells - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    99
    Thermo Fisher expression vector pcdna3 1
    Endothelial cell-derived exosomes mediate transmission of LGTV RNA from bEnd.3 cells to <t>N2a</t> neuronal cells. The bEnd.3 cells (1 x 10 5 ) were infected with 6 MOI of LGTV. QRT-PCR analysis showing total LGTV loads (A) or levels of LGTV positive-sense strand or negative-sense strand (B) in exosomes isolated from bEnd.3 cells at different time points (24, 48, 72, 96 and 120 h p.i.). (C) Viral loads were determined after 24 h p.i. by the presence of LGTV in N2a cells (1 x 10 5 ) treated with LGTV containing exosomes (20 μl) or supernatant (400 μl) fractions prepared from 24 or 48 h p.i. of bEnd.3 cells. (D) Viral loads at 48 h p.i. of N2a cells (plated in lower chamber) as determined by the presence of LGTV in a transwell assay performed with 1x 10 5 of bEnd.3 cells treated with 20 μl brain endothelial cell-derived exosomes (in upper chamber) for 4 h in the presence or absence of exosome inhibitor (5 μM) (in upper chamber) or infected with LGTV laboratory virus stocks is shown. LGTV transcript levels were normalized to mouse beta-actin. P value determined by Student’s two-tail t test is shown. Representative data is shown from three independent experiments.
    Expression Vector Pcdna3 1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2833 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/expression vector pcdna3 1/product/Thermo Fisher
    Average 99 stars, based on 2833 article reviews
    Price from $9.99 to $1999.99
    expression vector pcdna3 1 - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    90
    Sarstedt sterile screw cap tubes
    Endothelial cell-derived exosomes mediate transmission of LGTV RNA from bEnd.3 cells to <t>N2a</t> neuronal cells. The bEnd.3 cells (1 x 10 5 ) were infected with 6 MOI of LGTV. QRT-PCR analysis showing total LGTV loads (A) or levels of LGTV positive-sense strand or negative-sense strand (B) in exosomes isolated from bEnd.3 cells at different time points (24, 48, 72, 96 and 120 h p.i.). (C) Viral loads were determined after 24 h p.i. by the presence of LGTV in N2a cells (1 x 10 5 ) treated with LGTV containing exosomes (20 μl) or supernatant (400 μl) fractions prepared from 24 or 48 h p.i. of bEnd.3 cells. (D) Viral loads at 48 h p.i. of N2a cells (plated in lower chamber) as determined by the presence of LGTV in a transwell assay performed with 1x 10 5 of bEnd.3 cells treated with 20 μl brain endothelial cell-derived exosomes (in upper chamber) for 4 h in the presence or absence of exosome inhibitor (5 μM) (in upper chamber) or infected with LGTV laboratory virus stocks is shown. LGTV transcript levels were normalized to mouse beta-actin. P value determined by Student’s two-tail t test is shown. Representative data is shown from three independent experiments.
    Sterile Screw Cap Tubes, supplied by Sarstedt, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sterile screw cap tubes/product/Sarstedt
    Average 90 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    sterile screw cap tubes - by Bioz Stars, 2020-08
    90/100 stars
      Buy from Supplier

    89
    Promega pnl1 1
    Endothelial cell-derived exosomes mediate transmission of LGTV RNA from bEnd.3 cells to <t>N2a</t> neuronal cells. The bEnd.3 cells (1 x 10 5 ) were infected with 6 MOI of LGTV. QRT-PCR analysis showing total LGTV loads (A) or levels of LGTV positive-sense strand or negative-sense strand (B) in exosomes isolated from bEnd.3 cells at different time points (24, 48, 72, 96 and 120 h p.i.). (C) Viral loads were determined after 24 h p.i. by the presence of LGTV in N2a cells (1 x 10 5 ) treated with LGTV containing exosomes (20 μl) or supernatant (400 μl) fractions prepared from 24 or 48 h p.i. of bEnd.3 cells. (D) Viral loads at 48 h p.i. of N2a cells (plated in lower chamber) as determined by the presence of LGTV in a transwell assay performed with 1x 10 5 of bEnd.3 cells treated with 20 μl brain endothelial cell-derived exosomes (in upper chamber) for 4 h in the presence or absence of exosome inhibitor (5 μM) (in upper chamber) or infected with LGTV laboratory virus stocks is shown. LGTV transcript levels were normalized to mouse beta-actin. P value determined by Student’s two-tail t test is shown. Representative data is shown from three independent experiments.
    Pnl1 1, supplied by Promega, used in various techniques. Bioz Stars score: 89/100, based on 346 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pnl1 1/product/Promega
    Average 89 stars, based on 346 article reviews
    Price from $9.99 to $1999.99
    pnl1 1 - by Bioz Stars, 2020-08
    89/100 stars
      Buy from Supplier

    99
    New England Biolabs avr ii
    Endothelial cell-derived exosomes mediate transmission of LGTV RNA from bEnd.3 cells to <t>N2a</t> neuronal cells. The bEnd.3 cells (1 x 10 5 ) were infected with 6 MOI of LGTV. QRT-PCR analysis showing total LGTV loads (A) or levels of LGTV positive-sense strand or negative-sense strand (B) in exosomes isolated from bEnd.3 cells at different time points (24, 48, 72, 96 and 120 h p.i.). (C) Viral loads were determined after 24 h p.i. by the presence of LGTV in N2a cells (1 x 10 5 ) treated with LGTV containing exosomes (20 μl) or supernatant (400 μl) fractions prepared from 24 or 48 h p.i. of bEnd.3 cells. (D) Viral loads at 48 h p.i. of N2a cells (plated in lower chamber) as determined by the presence of LGTV in a transwell assay performed with 1x 10 5 of bEnd.3 cells treated with 20 μl brain endothelial cell-derived exosomes (in upper chamber) for 4 h in the presence or absence of exosome inhibitor (5 μM) (in upper chamber) or infected with LGTV laboratory virus stocks is shown. LGTV transcript levels were normalized to mouse beta-actin. P value determined by Student’s two-tail t test is shown. Representative data is shown from three independent experiments.
    Avr Ii, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 403 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/avr ii/product/New England Biolabs
    Average 99 stars, based on 403 article reviews
    Price from $9.99 to $1999.99
    avr ii - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    99
    Bio-Rad gene pulser xcell electroporation system
    Endothelial cell-derived exosomes mediate transmission of LGTV RNA from bEnd.3 cells to <t>N2a</t> neuronal cells. The bEnd.3 cells (1 x 10 5 ) were infected with 6 MOI of LGTV. QRT-PCR analysis showing total LGTV loads (A) or levels of LGTV positive-sense strand or negative-sense strand (B) in exosomes isolated from bEnd.3 cells at different time points (24, 48, 72, 96 and 120 h p.i.). (C) Viral loads were determined after 24 h p.i. by the presence of LGTV in N2a cells (1 x 10 5 ) treated with LGTV containing exosomes (20 μl) or supernatant (400 μl) fractions prepared from 24 or 48 h p.i. of bEnd.3 cells. (D) Viral loads at 48 h p.i. of N2a cells (plated in lower chamber) as determined by the presence of LGTV in a transwell assay performed with 1x 10 5 of bEnd.3 cells treated with 20 μl brain endothelial cell-derived exosomes (in upper chamber) for 4 h in the presence or absence of exosome inhibitor (5 μM) (in upper chamber) or infected with LGTV laboratory virus stocks is shown. LGTV transcript levels were normalized to mouse beta-actin. P value determined by Student’s two-tail t test is shown. Representative data is shown from three independent experiments.
    Gene Pulser Xcell Electroporation System, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 2856 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gene pulser xcell electroporation system/product/Bio-Rad
    Average 99 stars, based on 2856 article reviews
    Price from $9.99 to $1999.99
    gene pulser xcell electroporation system - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    99
    Qiagen endo free plasmid maxikit
    Endothelial cell-derived exosomes mediate transmission of LGTV RNA from bEnd.3 cells to <t>N2a</t> neuronal cells. The bEnd.3 cells (1 x 10 5 ) were infected with 6 MOI of LGTV. QRT-PCR analysis showing total LGTV loads (A) or levels of LGTV positive-sense strand or negative-sense strand (B) in exosomes isolated from bEnd.3 cells at different time points (24, 48, 72, 96 and 120 h p.i.). (C) Viral loads were determined after 24 h p.i. by the presence of LGTV in N2a cells (1 x 10 5 ) treated with LGTV containing exosomes (20 μl) or supernatant (400 μl) fractions prepared from 24 or 48 h p.i. of bEnd.3 cells. (D) Viral loads at 48 h p.i. of N2a cells (plated in lower chamber) as determined by the presence of LGTV in a transwell assay performed with 1x 10 5 of bEnd.3 cells treated with 20 μl brain endothelial cell-derived exosomes (in upper chamber) for 4 h in the presence or absence of exosome inhibitor (5 μM) (in upper chamber) or infected with LGTV laboratory virus stocks is shown. LGTV transcript levels were normalized to mouse beta-actin. P value determined by Student’s two-tail t test is shown. Representative data is shown from three independent experiments.
    Endo Free Plasmid Maxikit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/endo free plasmid maxikit/product/Qiagen
    Average 99 stars, based on 16 article reviews
    Price from $9.99 to $1999.99
    endo free plasmid maxikit - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    92
    Bio-Rad rna transcripts
    Endothelial cell-derived exosomes mediate transmission of LGTV RNA from bEnd.3 cells to <t>N2a</t> neuronal cells. The bEnd.3 cells (1 x 10 5 ) were infected with 6 MOI of LGTV. QRT-PCR analysis showing total LGTV loads (A) or levels of LGTV positive-sense strand or negative-sense strand (B) in exosomes isolated from bEnd.3 cells at different time points (24, 48, 72, 96 and 120 h p.i.). (C) Viral loads were determined after 24 h p.i. by the presence of LGTV in N2a cells (1 x 10 5 ) treated with LGTV containing exosomes (20 μl) or supernatant (400 μl) fractions prepared from 24 or 48 h p.i. of bEnd.3 cells. (D) Viral loads at 48 h p.i. of N2a cells (plated in lower chamber) as determined by the presence of LGTV in a transwell assay performed with 1x 10 5 of bEnd.3 cells treated with 20 μl brain endothelial cell-derived exosomes (in upper chamber) for 4 h in the presence or absence of exosome inhibitor (5 μM) (in upper chamber) or infected with LGTV laboratory virus stocks is shown. LGTV transcript levels were normalized to mouse beta-actin. P value determined by Student’s two-tail t test is shown. Representative data is shown from three independent experiments.
    Rna Transcripts, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 549 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rna transcripts/product/Bio-Rad
    Average 92 stars, based on 549 article reviews
    Price from $9.99 to $1999.99
    rna transcripts - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    99
    Thermo Fisher heat inactivated fbs
    Endothelial cell-derived exosomes mediate transmission of LGTV RNA from bEnd.3 cells to <t>N2a</t> neuronal cells. The bEnd.3 cells (1 x 10 5 ) were infected with 6 MOI of LGTV. QRT-PCR analysis showing total LGTV loads (A) or levels of LGTV positive-sense strand or negative-sense strand (B) in exosomes isolated from bEnd.3 cells at different time points (24, 48, 72, 96 and 120 h p.i.). (C) Viral loads were determined after 24 h p.i. by the presence of LGTV in N2a cells (1 x 10 5 ) treated with LGTV containing exosomes (20 μl) or supernatant (400 μl) fractions prepared from 24 or 48 h p.i. of bEnd.3 cells. (D) Viral loads at 48 h p.i. of N2a cells (plated in lower chamber) as determined by the presence of LGTV in a transwell assay performed with 1x 10 5 of bEnd.3 cells treated with 20 μl brain endothelial cell-derived exosomes (in upper chamber) for 4 h in the presence or absence of exosome inhibitor (5 μM) (in upper chamber) or infected with LGTV laboratory virus stocks is shown. LGTV transcript levels were normalized to mouse beta-actin. P value determined by Student’s two-tail t test is shown. Representative data is shown from three independent experiments.
    Heat Inactivated Fbs, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 10705 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/heat inactivated fbs/product/Thermo Fisher
    Average 99 stars, based on 10705 article reviews
    Price from $9.99 to $1999.99
    heat inactivated fbs - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    Image Search Results


    Analysis of Ago and Dcr protein-encoding genes in the Ixodes scapularis genome. ( A ) and ( B ) are gene trees for metazoan Ago-subfamily genes (A) and Dcr genes (B) respectively, constructed using a Bayesian approach under a GTR model (nodes are labeled if they receive > 90% support; see Materials and Methods). Trees are unrooted, but presented as if the root fell between the two Cnidarian homologs. ( C ) Detection of transcripts encoding Ago and Dcr proteins in IDE8 cells by RT-PCR. RNA not treated with reverse transcriptase was used as control during the PCR reaction (no RT). See also Supplementary Figure S5.

    Journal: Nucleic Acids Research

    Article Title: Induction and suppression of tick cell antiviral RNAi responses by tick-borne flaviviruses

    doi: 10.1093/nar/gku657

    Figure Lengend Snippet: Analysis of Ago and Dcr protein-encoding genes in the Ixodes scapularis genome. ( A ) and ( B ) are gene trees for metazoan Ago-subfamily genes (A) and Dcr genes (B) respectively, constructed using a Bayesian approach under a GTR model (nodes are labeled if they receive > 90% support; see Materials and Methods). Trees are unrooted, but presented as if the root fell between the two Cnidarian homologs. ( C ) Detection of transcripts encoding Ago and Dcr proteins in IDE8 cells by RT-PCR. RNA not treated with reverse transcriptase was used as control during the PCR reaction (no RT). See also Supplementary Figure S5.

    Article Snippet: Transcription of putative Ago and Dcr genes was verified in IDE8 cells (Figure ).

    Techniques: Construct, Labeling, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction

    Analysis of subgenomic flavivirus (sf)RNA in the 3′UTR of tick-borne flaviviruses. ( A ) Structure model of SL 2 and SL 1 RNA stem loop structures of TBEV and LGTV. ( B ) Expression of TBEV (TND/ΔME) (top) and LGTV (bottom) sfRNA in replicon (top), non-transfected (control CTRL) or infected cells (bottom) was detected by northern blot analysis with 3′UTR specific DIG-PCR probes. ( C ) The effect of sfRNA on RNAi in IDE8 cells was determined by co-transfection of FFluc , Rluc and expression constructs for MBP-HdVr (MBP), LGTV 3′UTR or TBEV 3′UTR. Silencing was induced 24 hpt following addition of dsRNA to the culture medium. At 48 hpt, relative luciferase activity ( FFluc /Rluc) was determined and normalized to cells treated with eGFP specific (ctrl) dsRNA. The luciferase expression level measured with MBP-HdVr was set at 1.0. The mean with standard error is shown for three independent experiments performed in duplicate (one experiment)/triplicate (two experiments). * indicate significance by Tukey's HSD ( P ≤ 0.05). See also Supplementary Figure S7.

    Journal: Nucleic Acids Research

    Article Title: Induction and suppression of tick cell antiviral RNAi responses by tick-borne flaviviruses

    doi: 10.1093/nar/gku657

    Figure Lengend Snippet: Analysis of subgenomic flavivirus (sf)RNA in the 3′UTR of tick-borne flaviviruses. ( A ) Structure model of SL 2 and SL 1 RNA stem loop structures of TBEV and LGTV. ( B ) Expression of TBEV (TND/ΔME) (top) and LGTV (bottom) sfRNA in replicon (top), non-transfected (control CTRL) or infected cells (bottom) was detected by northern blot analysis with 3′UTR specific DIG-PCR probes. ( C ) The effect of sfRNA on RNAi in IDE8 cells was determined by co-transfection of FFluc , Rluc and expression constructs for MBP-HdVr (MBP), LGTV 3′UTR or TBEV 3′UTR. Silencing was induced 24 hpt following addition of dsRNA to the culture medium. At 48 hpt, relative luciferase activity ( FFluc /Rluc) was determined and normalized to cells treated with eGFP specific (ctrl) dsRNA. The luciferase expression level measured with MBP-HdVr was set at 1.0. The mean with standard error is shown for three independent experiments performed in duplicate (one experiment)/triplicate (two experiments). * indicate significance by Tukey's HSD ( P ≤ 0.05). See also Supplementary Figure S7.

    Article Snippet: Transcription of putative Ago and Dcr genes was verified in IDE8 cells (Figure ).

    Techniques: Expressing, Transfection, Infection, Northern Blot, Polymerase Chain Reaction, Cotransfection, Construct, Luciferase, Activity Assay

    Characterization of exogenous-derived small RNAs in IDE8 cells. ( A ) Size distribution of small RNA molecules mapping either to LGTV E5repRluc2B/3 replicon (left panel) at 48 hpt or LGTV TP21 (right panel) at 72 hpi in IDE8 cells. ( B ) Frequency distribution of 22 nt small RNA molecules mapped to the E5repRluc 2B/3 replicon (5′UTR to 3′UTR) (left panel) or LGTV TP21 (right panel). The y-axis shows the frequency of the 22 nt siRNAs mapping to the corresponding nucleotide position in the x-axis. Positive numbers and dark gray peaks represent the frequency of siRNAs mapping to the genome (in 5′-3′ orientation) and light gray peaks/negative numbers to the antigenome (in 3′-5′ orientation). See also Supplementary Figure S2. ( C ) Frequency map of 22 nt small RNAs mapping to the opposite strand of the LGTV replicon (left panel) or LGTV TP21 (right panel).

    Journal: Nucleic Acids Research

    Article Title: Induction and suppression of tick cell antiviral RNAi responses by tick-borne flaviviruses

    doi: 10.1093/nar/gku657

    Figure Lengend Snippet: Characterization of exogenous-derived small RNAs in IDE8 cells. ( A ) Size distribution of small RNA molecules mapping either to LGTV E5repRluc2B/3 replicon (left panel) at 48 hpt or LGTV TP21 (right panel) at 72 hpi in IDE8 cells. ( B ) Frequency distribution of 22 nt small RNA molecules mapped to the E5repRluc 2B/3 replicon (5′UTR to 3′UTR) (left panel) or LGTV TP21 (right panel). The y-axis shows the frequency of the 22 nt siRNAs mapping to the corresponding nucleotide position in the x-axis. Positive numbers and dark gray peaks represent the frequency of siRNAs mapping to the genome (in 5′-3′ orientation) and light gray peaks/negative numbers to the antigenome (in 3′-5′ orientation). See also Supplementary Figure S2. ( C ) Frequency map of 22 nt small RNAs mapping to the opposite strand of the LGTV replicon (left panel) or LGTV TP21 (right panel).

    Article Snippet: Transcription of putative Ago and Dcr genes was verified in IDE8 cells (Figure ).

    Techniques: Derivative Assay

    Knockdown of transcripts encoding Ago or Dcr proteins. ( A ) dsRNA-based silencing of Ago and Dcr encoding transcripts in IDE8 cells. Transcripts were detected by RT-PCR using gene-specific primers. A PCR product of 16S ribosomal RNA was used as housekeeping gene and eGFP specific dsRNA treated cells as control (crtl). ( B ) mRNA knockdowns quantification by Image J software, using 16S as control. The relative mean (normalized to eGFP-dsRNA controls) with standard error is shown for at least 10 repeats.

    Journal: Nucleic Acids Research

    Article Title: Induction and suppression of tick cell antiviral RNAi responses by tick-borne flaviviruses

    doi: 10.1093/nar/gku657

    Figure Lengend Snippet: Knockdown of transcripts encoding Ago or Dcr proteins. ( A ) dsRNA-based silencing of Ago and Dcr encoding transcripts in IDE8 cells. Transcripts were detected by RT-PCR using gene-specific primers. A PCR product of 16S ribosomal RNA was used as housekeeping gene and eGFP specific dsRNA treated cells as control (crtl). ( B ) mRNA knockdowns quantification by Image J software, using 16S as control. The relative mean (normalized to eGFP-dsRNA controls) with standard error is shown for at least 10 repeats.

    Article Snippet: Transcription of putative Ago and Dcr genes was verified in IDE8 cells (Figure ).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Software

    Endothelial cell-derived exosomes mediate transmission of LGTV RNA from bEnd.3 cells to N2a neuronal cells. The bEnd.3 cells (1 x 10 5 ) were infected with 6 MOI of LGTV. QRT-PCR analysis showing total LGTV loads (A) or levels of LGTV positive-sense strand or negative-sense strand (B) in exosomes isolated from bEnd.3 cells at different time points (24, 48, 72, 96 and 120 h p.i.). (C) Viral loads were determined after 24 h p.i. by the presence of LGTV in N2a cells (1 x 10 5 ) treated with LGTV containing exosomes (20 μl) or supernatant (400 μl) fractions prepared from 24 or 48 h p.i. of bEnd.3 cells. (D) Viral loads at 48 h p.i. of N2a cells (plated in lower chamber) as determined by the presence of LGTV in a transwell assay performed with 1x 10 5 of bEnd.3 cells treated with 20 μl brain endothelial cell-derived exosomes (in upper chamber) for 4 h in the presence or absence of exosome inhibitor (5 μM) (in upper chamber) or infected with LGTV laboratory virus stocks is shown. LGTV transcript levels were normalized to mouse beta-actin. P value determined by Student’s two-tail t test is shown. Representative data is shown from three independent experiments.

    Journal: PLoS Pathogens

    Article Title: Exosomes serve as novel modes of tick-borne flavivirus transmission from arthropod to human cells and facilitates dissemination of viral RNA and proteins to the vertebrate neuronal cells

    doi: 10.1371/journal.ppat.1006764

    Figure Lengend Snippet: Endothelial cell-derived exosomes mediate transmission of LGTV RNA from bEnd.3 cells to N2a neuronal cells. The bEnd.3 cells (1 x 10 5 ) were infected with 6 MOI of LGTV. QRT-PCR analysis showing total LGTV loads (A) or levels of LGTV positive-sense strand or negative-sense strand (B) in exosomes isolated from bEnd.3 cells at different time points (24, 48, 72, 96 and 120 h p.i.). (C) Viral loads were determined after 24 h p.i. by the presence of LGTV in N2a cells (1 x 10 5 ) treated with LGTV containing exosomes (20 μl) or supernatant (400 μl) fractions prepared from 24 or 48 h p.i. of bEnd.3 cells. (D) Viral loads at 48 h p.i. of N2a cells (plated in lower chamber) as determined by the presence of LGTV in a transwell assay performed with 1x 10 5 of bEnd.3 cells treated with 20 μl brain endothelial cell-derived exosomes (in upper chamber) for 4 h in the presence or absence of exosome inhibitor (5 μM) (in upper chamber) or infected with LGTV laboratory virus stocks is shown. LGTV transcript levels were normalized to mouse beta-actin. P value determined by Student’s two-tail t test is shown. Representative data is shown from three independent experiments.

    Article Snippet: Exosome fractions containing unknown PFU (plaque forming units) of LGTV viral genomes were collected from tick cells (5 x 106 cells) or N2a cells or murine cortical neurons (1 x 105 cells) and resuspended in 250 μl of PBS, 30 μl of this suspension (exosome fraction) was used for plaque assays.

    Techniques: Derivative Assay, Transmission Assay, Infection, Quantitative RT-PCR, Isolation, Transwell Assay

    Transmission of LGTV through infectious exosome to naïve cells is clathrin dependent. (A) Quantitative assessment of number of plaques from exosomal and supernatant fractions is shown. TMTC indicates “too many to count”. (B) QRT-PCR analysis for the LGTV loads in fresh N2a (1 x 10 5 ) cells at 24 h p.i., infected with exosomes (20 μl) or supernatant (400 μl) fractions prepared from 24, 48 and 72 h p.i. (C) Mouse N2a cells showing infection kinetics with WNV (MOI 5) at different times (24, 48, 72 h) p.i., WNV E gene transcript levels were normalized to mouse beta-actin. (D) Relative loads of WNV in infected N2a cell-derived exosomes collected at 24, 48, 72 h p.i. LGTV loads (72 h p.i.) in N2a cells treated with either 4G2 antibody (5 μg for 4 h) (E) or Pitstop-2 clathrin inhibitor (F) (treated 30 μM for 15 min at 37°C) and infected with LGTV-containing (72 h p.i.) N2a cell-derived exosomes is shown. Infected but untreated samples serve as control in (E). Infected but DMSO-treated serve as control in (F). LGTV transcript levels were normalized to mouse beta-actin. P value determined by Student’s two-tail t test is shown. Representative data is shown from at least three independent experiments.

    Journal: PLoS Pathogens

    Article Title: Exosomes serve as novel modes of tick-borne flavivirus transmission from arthropod to human cells and facilitates dissemination of viral RNA and proteins to the vertebrate neuronal cells

    doi: 10.1371/journal.ppat.1006764

    Figure Lengend Snippet: Transmission of LGTV through infectious exosome to naïve cells is clathrin dependent. (A) Quantitative assessment of number of plaques from exosomal and supernatant fractions is shown. TMTC indicates “too many to count”. (B) QRT-PCR analysis for the LGTV loads in fresh N2a (1 x 10 5 ) cells at 24 h p.i., infected with exosomes (20 μl) or supernatant (400 μl) fractions prepared from 24, 48 and 72 h p.i. (C) Mouse N2a cells showing infection kinetics with WNV (MOI 5) at different times (24, 48, 72 h) p.i., WNV E gene transcript levels were normalized to mouse beta-actin. (D) Relative loads of WNV in infected N2a cell-derived exosomes collected at 24, 48, 72 h p.i. LGTV loads (72 h p.i.) in N2a cells treated with either 4G2 antibody (5 μg for 4 h) (E) or Pitstop-2 clathrin inhibitor (F) (treated 30 μM for 15 min at 37°C) and infected with LGTV-containing (72 h p.i.) N2a cell-derived exosomes is shown. Infected but untreated samples serve as control in (E). Infected but DMSO-treated serve as control in (F). LGTV transcript levels were normalized to mouse beta-actin. P value determined by Student’s two-tail t test is shown. Representative data is shown from at least three independent experiments.

    Article Snippet: Exosome fractions containing unknown PFU (plaque forming units) of LGTV viral genomes were collected from tick cells (5 x 106 cells) or N2a cells or murine cortical neurons (1 x 105 cells) and resuspended in 250 μl of PBS, 30 μl of this suspension (exosome fraction) was used for plaque assays.

    Techniques: Transmission Assay, Quantitative RT-PCR, Infection, Derivative Assay

    Detection of LGTV RNA and proteins in exosomes isolated from N2a neuronal cell line. (A) Cryo-EM images showing exosomes isolated from uninfected and LGTV-infected (MOI 6; 72 h p.i.), N2a cells (1 x 10 7 ). Scale indicates 100nm. Quantification of diameter or sizes for heterogenous population of exosomes from uninfected (B) or LGTV- infected (C) N2a cell-derived exosomes. Number of exosomes analyzed were n = 32 (uninfected) and n = 131 (infected) groups. (D) Comparison of exosome numbers per image from uninfected (n = 9) and infected (n = 13) groups is shown. (E) DG-Exos showing presence of enhanced LGTV envelope [E]-protein loads in fractions 1–6. Fractions 3–5 showed enriched amounts of exosomal markers CD9 and HSP70. E-protein detection in fraction 5 in 0.1 μm filtered samples processed for OptiPrep DG-isolation is shown. QRT-PCR analysis showing levels of total LGTV loads (F), copy numbers (G) and LGTV positive-sense strand or negative-sense strand (H) in exosomes isolated from N2a cells at different time points. N2a (1 x 10 5 ) cells were infected with 6 MOI of LGTV, and LGTV loads were analyzed at 72 h p.i. (I) Treatment of LGTV-infected (72 h p.i.) N2a cell-derived exosomes with RNase A is shown. The uninfected samples treated with RNase serve as control. LGTV transcript levels were normalized to mouse beta-actin. (J) Immunoblotting analysis showing detection of LGTV E glycoprotein and mammalian exosomal marker CD9 in exosome fractions and total lysates from whole cells prepared at 48 h p.i. from 2 x 10 6 uninfected (UI) or infected (I) N2a cells. Stain-free gels showing total protein profiles serve as the loading control. (K) Native-PAGE followed by immunoblotting analysis showing presence of LGTV E- and NS1 proteins from LGTV-infected (MOI 1; 72 h p.i.) or uninfected N2a cell-derived exosomes treated with Triton-X-100 (0.03%), or freeze-thaw cycles (3 times freezing at -80°C for 1 h each cycle) or untreated, held on ice. Coomassie stained gel image showing total protein profiles serve as a loading control. (L) ELISA performed on uninfected or LGTV-infected (MOI 6; 72 h p.i.) N2a cell-derived exosomes either untreated or treated with Triton-X-100 (0.1%). (M) Antibody-beads binding assay performed on LGTV infected (MOI 1, 72 h p.i.) N2a cell-derived exosomes (collected from N2a cells either untreated or treated with GW4869; 5 μM, exosome inhibitor) showing no differences in LGTV-E protein loads between 4G2 or isotype and untreated samples.

    Journal: PLoS Pathogens

    Article Title: Exosomes serve as novel modes of tick-borne flavivirus transmission from arthropod to human cells and facilitates dissemination of viral RNA and proteins to the vertebrate neuronal cells

    doi: 10.1371/journal.ppat.1006764

    Figure Lengend Snippet: Detection of LGTV RNA and proteins in exosomes isolated from N2a neuronal cell line. (A) Cryo-EM images showing exosomes isolated from uninfected and LGTV-infected (MOI 6; 72 h p.i.), N2a cells (1 x 10 7 ). Scale indicates 100nm. Quantification of diameter or sizes for heterogenous population of exosomes from uninfected (B) or LGTV- infected (C) N2a cell-derived exosomes. Number of exosomes analyzed were n = 32 (uninfected) and n = 131 (infected) groups. (D) Comparison of exosome numbers per image from uninfected (n = 9) and infected (n = 13) groups is shown. (E) DG-Exos showing presence of enhanced LGTV envelope [E]-protein loads in fractions 1–6. Fractions 3–5 showed enriched amounts of exosomal markers CD9 and HSP70. E-protein detection in fraction 5 in 0.1 μm filtered samples processed for OptiPrep DG-isolation is shown. QRT-PCR analysis showing levels of total LGTV loads (F), copy numbers (G) and LGTV positive-sense strand or negative-sense strand (H) in exosomes isolated from N2a cells at different time points. N2a (1 x 10 5 ) cells were infected with 6 MOI of LGTV, and LGTV loads were analyzed at 72 h p.i. (I) Treatment of LGTV-infected (72 h p.i.) N2a cell-derived exosomes with RNase A is shown. The uninfected samples treated with RNase serve as control. LGTV transcript levels were normalized to mouse beta-actin. (J) Immunoblotting analysis showing detection of LGTV E glycoprotein and mammalian exosomal marker CD9 in exosome fractions and total lysates from whole cells prepared at 48 h p.i. from 2 x 10 6 uninfected (UI) or infected (I) N2a cells. Stain-free gels showing total protein profiles serve as the loading control. (K) Native-PAGE followed by immunoblotting analysis showing presence of LGTV E- and NS1 proteins from LGTV-infected (MOI 1; 72 h p.i.) or uninfected N2a cell-derived exosomes treated with Triton-X-100 (0.03%), or freeze-thaw cycles (3 times freezing at -80°C for 1 h each cycle) or untreated, held on ice. Coomassie stained gel image showing total protein profiles serve as a loading control. (L) ELISA performed on uninfected or LGTV-infected (MOI 6; 72 h p.i.) N2a cell-derived exosomes either untreated or treated with Triton-X-100 (0.1%). (M) Antibody-beads binding assay performed on LGTV infected (MOI 1, 72 h p.i.) N2a cell-derived exosomes (collected from N2a cells either untreated or treated with GW4869; 5 μM, exosome inhibitor) showing no differences in LGTV-E protein loads between 4G2 or isotype and untreated samples.

    Article Snippet: Exosome fractions containing unknown PFU (plaque forming units) of LGTV viral genomes were collected from tick cells (5 x 106 cells) or N2a cells or murine cortical neurons (1 x 105 cells) and resuspended in 250 μl of PBS, 30 μl of this suspension (exosome fraction) was used for plaque assays.

    Techniques: Isolation, Infection, Derivative Assay, Quantitative RT-PCR, Marker, Staining, Clear Native PAGE, Enzyme-linked Immunosorbent Assay, Binding Assay

    Treatment of neuronal N2a cells with exosome inhibitor affects LGTV infection, if treated either before or after infection. (A) QRT-PCR analysis showing levels of LGTV in exosomes isolated from N2a cells at 48 h p.i. in the presence of exosomes-inhibitor at different concentrations (1, 5, 10 μM). 1 x 10 5 N2a cells were pre-treated with inhibitor for 4 h followed by infection with LGTV (6 MOI). Exosomes isolated from N2a cells treated with DMSO serve as loading control. (B) Levels of LGTV in N2a cells at 48 h p.i. infected by treatment with exosomes (20 μl) isolated from control or inhibitor-treated N2a cells is shown. UI indicates uninfected cells that serve as control. (C) QRT-PCR analysis showing levels of LGTV in N2a cells at 72 h p.i. Uninfected (UI) N2a cells were treated with 5 μM inhibitor. For LGTV-infection, N2a cells were first treated for 4 h with inhibitor then followed by infection (Inhibitor/I) or cells were first infected for 4 h with LGTV and then treated with inhibitor (I/inhibitor). LGTV (6 MOI) was used to infect 1 x 10 5 N2a cells in the presence (5 μM) or absence of inhibitor. LGTV transcript levels were normalized to mouse beta-actin. (D) Plaque assays p erformed with different dilutions (1:10, 1:100, 1:1000) of exosomes fraction isolated from control DMSO-treated or inhibitor-treated N2a cells is shown. A representative image from two independent experiments is shown. (E) Quantitative assessment of number of plaques from DMSO-treated or inhibitor-treated (D) is shown. P value determined by Student’s two-tail t test is shown. (F) Laboratory virus stocks treated with either GW4869 inhibitor (5 and 10 μM) or DMSO as control is shown. Representative data in other panels is shown from at least three independent experiments.

    Journal: PLoS Pathogens

    Article Title: Exosomes serve as novel modes of tick-borne flavivirus transmission from arthropod to human cells and facilitates dissemination of viral RNA and proteins to the vertebrate neuronal cells

    doi: 10.1371/journal.ppat.1006764

    Figure Lengend Snippet: Treatment of neuronal N2a cells with exosome inhibitor affects LGTV infection, if treated either before or after infection. (A) QRT-PCR analysis showing levels of LGTV in exosomes isolated from N2a cells at 48 h p.i. in the presence of exosomes-inhibitor at different concentrations (1, 5, 10 μM). 1 x 10 5 N2a cells were pre-treated with inhibitor for 4 h followed by infection with LGTV (6 MOI). Exosomes isolated from N2a cells treated with DMSO serve as loading control. (B) Levels of LGTV in N2a cells at 48 h p.i. infected by treatment with exosomes (20 μl) isolated from control or inhibitor-treated N2a cells is shown. UI indicates uninfected cells that serve as control. (C) QRT-PCR analysis showing levels of LGTV in N2a cells at 72 h p.i. Uninfected (UI) N2a cells were treated with 5 μM inhibitor. For LGTV-infection, N2a cells were first treated for 4 h with inhibitor then followed by infection (Inhibitor/I) or cells were first infected for 4 h with LGTV and then treated with inhibitor (I/inhibitor). LGTV (6 MOI) was used to infect 1 x 10 5 N2a cells in the presence (5 μM) or absence of inhibitor. LGTV transcript levels were normalized to mouse beta-actin. (D) Plaque assays p erformed with different dilutions (1:10, 1:100, 1:1000) of exosomes fraction isolated from control DMSO-treated or inhibitor-treated N2a cells is shown. A representative image from two independent experiments is shown. (E) Quantitative assessment of number of plaques from DMSO-treated or inhibitor-treated (D) is shown. P value determined by Student’s two-tail t test is shown. (F) Laboratory virus stocks treated with either GW4869 inhibitor (5 and 10 μM) or DMSO as control is shown. Representative data in other panels is shown from at least three independent experiments.

    Article Snippet: Exosome fractions containing unknown PFU (plaque forming units) of LGTV viral genomes were collected from tick cells (5 x 106 cells) or N2a cells or murine cortical neurons (1 x 105 cells) and resuspended in 250 μl of PBS, 30 μl of this suspension (exosome fraction) was used for plaque assays.

    Techniques: Infection, Quantitative RT-PCR, Isolation