human lymphoblastoid cdna library Search Results


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  • 99
    New England Biolabs rrna depletion kit
    Rrna Depletion Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 582 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher total rna
    Total <t>RNA</t> sample data from a <t>lymphoblastoid</t> cell-line. All data are normalized against let-7a data. Relative signal intensity of the let-7 family conventional ( a ) and ProDeG ( b ) probes are shown after Cy-3 labeled total RNA from a lymphoblastoid cell-line was hybridized at 34°C. The relative amount of each let-7 family miRNA was quantified using TaqMan® qRT-PCR assay ( c ).
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    Nugen ovation human ffpe rna seq kit
    Total <t>RNA</t> sample data from a <t>lymphoblastoid</t> cell-line. All data are normalized against let-7a data. Relative signal intensity of the let-7 family conventional ( a ) and ProDeG ( b ) probes are shown after Cy-3 labeled total RNA from a lymphoblastoid cell-line was hybridized at 34°C. The relative amount of each let-7 family miRNA was quantified using TaqMan® qRT-PCR assay ( c ).
    Ovation Human Ffpe Rna Seq Kit, supplied by Nugen, used in various techniques. Bioz Stars score: 92/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen genomic dna
    Total <t>RNA</t> sample data from a <t>lymphoblastoid</t> cell-line. All data are normalized against let-7a data. Relative signal intensity of the let-7 family conventional ( a ) and ProDeG ( b ) probes are shown after Cy-3 labeled total RNA from a lymphoblastoid cell-line was hybridized at 34°C. The relative amount of each let-7 family miRNA was quantified using TaqMan® qRT-PCR assay ( c ).
    Genomic Dna, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 127055 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher vector pcr3
    Total <t>RNA</t> sample data from a <t>lymphoblastoid</t> cell-line. All data are normalized against let-7a data. Relative signal intensity of the let-7 family conventional ( a ) and ProDeG ( b ) probes are shown after Cy-3 labeled total RNA from a lymphoblastoid cell-line was hybridized at 34°C. The relative amount of each let-7 family miRNA was quantified using TaqMan® qRT-PCR assay ( c ).
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    Thermo Fisher ion proton system
    Total <t>RNA</t> sample data from a <t>lymphoblastoid</t> cell-line. All data are normalized against let-7a data. Relative signal intensity of the let-7 family conventional ( a ) and ProDeG ( b ) probes are shown after Cy-3 labeled total RNA from a lymphoblastoid cell-line was hybridized at 34°C. The relative amount of each let-7 family miRNA was quantified using TaqMan® qRT-PCR assay ( c ).
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    Thermo Fisher ion pi chip kit v3
    Total <t>RNA</t> sample data from a <t>lymphoblastoid</t> cell-line. All data are normalized against let-7a data. Relative signal intensity of the let-7 family conventional ( a ) and ProDeG ( b ) probes are shown after Cy-3 labeled total RNA from a lymphoblastoid cell-line was hybridized at 34°C. The relative amount of each let-7 family miRNA was quantified using TaqMan® qRT-PCR assay ( c ).
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    Thermo Fisher ion pi hi q sequencing 200 kit
    Total <t>RNA</t> sample data from a <t>lymphoblastoid</t> cell-line. All data are normalized against let-7a data. Relative signal intensity of the let-7 family conventional ( a ) and ProDeG ( b ) probes are shown after Cy-3 labeled total RNA from a lymphoblastoid cell-line was hybridized at 34°C. The relative amount of each let-7 family miRNA was quantified using TaqMan® qRT-PCR assay ( c ).
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    Santa Cruz Biotechnology scrambled non targeting sirna
    Cellular <t>KAP1</t> regulates EBV lytic cycle in Burkitt Lymphoma cells. A-C. HH514-16 Burkitt lymphoma (BL) cells were transfected with scrambled <t>siRNA</t> (non-targeting control; open bars in B, C) or siRNA to KAP1 (black bars in B, C); D-F. BL cells were transfected with empty vector (open bars in E, F) or pFLAG-CMV2-KAP1 (black bars in E, F). Transfected cells were treated with NaB 24 hours after transfection, and harvested at indicated times (A, D), 24 hours (B, E), or 48 hours (C, F) post-treatment to determine relative amounts of ZEBRA, KAP1 and β-actin by immunoblotting (A, D), relative levels of transcripts from EBV lytic genes BZLF1 , BMRF1 and BFRF3 by qRT-PCR after normalization to 18S rRNA using the ΔΔC T method (B, E), and relative levels of cell-associated EBV DNA by qPCR (C, F). Error bars: SEM of 3 experiments with 4 technical replicates. Western blots are representative of 2 experiments. Numbers below blots indicate relative amounts of protein after normalization to β-actin.
    Scrambled Non Targeting Sirna, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs nebnext ultra rna library prep kit for illumina
    Cellular <t>KAP1</t> regulates EBV lytic cycle in Burkitt Lymphoma cells. A-C. HH514-16 Burkitt lymphoma (BL) cells were transfected with scrambled <t>siRNA</t> (non-targeting control; open bars in B, C) or siRNA to KAP1 (black bars in B, C); D-F. BL cells were transfected with empty vector (open bars in E, F) or pFLAG-CMV2-KAP1 (black bars in E, F). Transfected cells were treated with NaB 24 hours after transfection, and harvested at indicated times (A, D), 24 hours (B, E), or 48 hours (C, F) post-treatment to determine relative amounts of ZEBRA, KAP1 and β-actin by immunoblotting (A, D), relative levels of transcripts from EBV lytic genes BZLF1 , BMRF1 and BFRF3 by qRT-PCR after normalization to 18S rRNA using the ΔΔC T method (B, E), and relative levels of cell-associated EBV DNA by qPCR (C, F). Error bars: SEM of 3 experiments with 4 technical replicates. Western blots are representative of 2 experiments. Numbers below blots indicate relative amounts of protein after normalization to β-actin.
    Nebnext Ultra Rna Library Prep Kit For Illumina, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 77 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher expression vector pcdna3 1
    Cellular <t>KAP1</t> regulates EBV lytic cycle in Burkitt Lymphoma cells. A-C. HH514-16 Burkitt lymphoma (BL) cells were transfected with scrambled <t>siRNA</t> (non-targeting control; open bars in B, C) or siRNA to KAP1 (black bars in B, C); D-F. BL cells were transfected with empty vector (open bars in E, F) or pFLAG-CMV2-KAP1 (black bars in E, F). Transfected cells were treated with NaB 24 hours after transfection, and harvested at indicated times (A, D), 24 hours (B, E), or 48 hours (C, F) post-treatment to determine relative amounts of ZEBRA, KAP1 and β-actin by immunoblotting (A, D), relative levels of transcripts from EBV lytic genes BZLF1 , BMRF1 and BFRF3 by qRT-PCR after normalization to 18S rRNA using the ΔΔC T method (B, E), and relative levels of cell-associated EBV DNA by qPCR (C, F). Error bars: SEM of 3 experiments with 4 technical replicates. Western blots are representative of 2 experiments. Numbers below blots indicate relative amounts of protein after normalization to β-actin.
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    5 PRIME rt pcr
    Cellular <t>KAP1</t> regulates EBV lytic cycle in Burkitt Lymphoma cells. A-C. HH514-16 Burkitt lymphoma (BL) cells were transfected with scrambled <t>siRNA</t> (non-targeting control; open bars in B, C) or siRNA to KAP1 (black bars in B, C); D-F. BL cells were transfected with empty vector (open bars in E, F) or pFLAG-CMV2-KAP1 (black bars in E, F). Transfected cells were treated with NaB 24 hours after transfection, and harvested at indicated times (A, D), 24 hours (B, E), or 48 hours (C, F) post-treatment to determine relative amounts of ZEBRA, KAP1 and β-actin by immunoblotting (A, D), relative levels of transcripts from EBV lytic genes BZLF1 , BMRF1 and BFRF3 by qRT-PCR after normalization to 18S rRNA using the ΔΔC T method (B, E), and relative levels of cell-associated EBV DNA by qPCR (C, F). Error bars: SEM of 3 experiments with 4 technical replicates. Western blots are representative of 2 experiments. Numbers below blots indicate relative amounts of protein after normalization to β-actin.
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    Thermo Fisher topo ta cloning kit
    Cellular <t>KAP1</t> regulates EBV lytic cycle in Burkitt Lymphoma cells. A-C. HH514-16 Burkitt lymphoma (BL) cells were transfected with scrambled <t>siRNA</t> (non-targeting control; open bars in B, C) or siRNA to KAP1 (black bars in B, C); D-F. BL cells were transfected with empty vector (open bars in E, F) or pFLAG-CMV2-KAP1 (black bars in E, F). Transfected cells were treated with NaB 24 hours after transfection, and harvested at indicated times (A, D), 24 hours (B, E), or 48 hours (C, F) post-treatment to determine relative amounts of ZEBRA, KAP1 and β-actin by immunoblotting (A, D), relative levels of transcripts from EBV lytic genes BZLF1 , BMRF1 and BFRF3 by qRT-PCR after normalization to 18S rRNA using the ΔΔC T method (B, E), and relative levels of cell-associated EBV DNA by qPCR (C, F). Error bars: SEM of 3 experiments with 4 technical replicates. Western blots are representative of 2 experiments. Numbers below blots indicate relative amounts of protein after normalization to β-actin.
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    Millipore ch7c17
    MVBs in T cells translocate to the IS. ( a ) J77 T cells were conjugated with SEE-pulsed or non-pulsed Raji B cells loaded with CMAC (blue). After 30 min, cells were fixed and stained for the MVB marker Hrs (red), CD3 and actin (both green). Plates show maximal projections of confocal images and the DIC images. ( b ) <t>CH7C17</t> T cells were conjugated with HA-loaded or non-loaded HOM2 B cells (blue). After 30 min, cells were fixed and stained for CD63 (green), CD3 and actin (both red). Plates show maximal projections of confocal images and the DIC images, scale bars applies to a and b . ( c ) Percentage of T and APC cells in which CD63, Hrs and CD3 relocalized to the T cell–APC contact area in the presence of HA peptide (filled bars) or its absence (open bars). Data are the arithmetic means±s.e.m. of four experiments. * P
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    Illumina Inc complementary dna libraries
    MVBs in T cells translocate to the IS. ( a ) J77 T cells were conjugated with SEE-pulsed or non-pulsed Raji B cells loaded with CMAC (blue). After 30 min, cells were fixed and stained for the MVB marker Hrs (red), CD3 and actin (both green). Plates show maximal projections of confocal images and the DIC images. ( b ) <t>CH7C17</t> T cells were conjugated with HA-loaded or non-loaded HOM2 B cells (blue). After 30 min, cells were fixed and stained for CD63 (green), CD3 and actin (both red). Plates show maximal projections of confocal images and the DIC images, scale bars applies to a and b . ( c ) Percentage of T and APC cells in which CD63, Hrs and CD3 relocalized to the T cell–APC contact area in the presence of HA peptide (filled bars) or its absence (open bars). Data are the arithmetic means±s.e.m. of four experiments. * P
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    Thermo Fisher control human brain tissues
    MVBs in T cells translocate to the IS. ( a ) J77 T cells were conjugated with SEE-pulsed or non-pulsed Raji B cells loaded with CMAC (blue). After 30 min, cells were fixed and stained for the MVB marker Hrs (red), CD3 and actin (both green). Plates show maximal projections of confocal images and the DIC images. ( b ) <t>CH7C17</t> T cells were conjugated with HA-loaded or non-loaded HOM2 B cells (blue). After 30 min, cells were fixed and stained for CD63 (green), CD3 and actin (both red). Plates show maximal projections of confocal images and the DIC images, scale bars applies to a and b . ( c ) Percentage of T and APC cells in which CD63, Hrs and CD3 relocalized to the T cell–APC contact area in the presence of HA peptide (filled bars) or its absence (open bars). Data are the arithmetic means±s.e.m. of four experiments. * P
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    Thermo Fisher human cot 1 dna
    MVBs in T cells translocate to the IS. ( a ) J77 T cells were conjugated with SEE-pulsed or non-pulsed Raji B cells loaded with CMAC (blue). After 30 min, cells were fixed and stained for the MVB marker Hrs (red), CD3 and actin (both green). Plates show maximal projections of confocal images and the DIC images. ( b ) <t>CH7C17</t> T cells were conjugated with HA-loaded or non-loaded HOM2 B cells (blue). After 30 min, cells were fixed and stained for CD63 (green), CD3 and actin (both red). Plates show maximal projections of confocal images and the DIC images, scale bars applies to a and b . ( c ) Percentage of T and APC cells in which CD63, Hrs and CD3 relocalized to the T cell–APC contact area in the presence of HA peptide (filled bars) or its absence (open bars). Data are the arithmetic means±s.e.m. of four experiments. * P
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    TaKaRa human brain regions
    MVBs in T cells translocate to the IS. ( a ) J77 T cells were conjugated with SEE-pulsed or non-pulsed Raji B cells loaded with CMAC (blue). After 30 min, cells were fixed and stained for the MVB marker Hrs (red), CD3 and actin (both green). Plates show maximal projections of confocal images and the DIC images. ( b ) <t>CH7C17</t> T cells were conjugated with HA-loaded or non-loaded HOM2 B cells (blue). After 30 min, cells were fixed and stained for CD63 (green), CD3 and actin (both red). Plates show maximal projections of confocal images and the DIC images, scale bars applies to a and b . ( c ) Percentage of T and APC cells in which CD63, Hrs and CD3 relocalized to the T cell–APC contact area in the presence of HA peptide (filled bars) or its absence (open bars). Data are the arithmetic means±s.e.m. of four experiments. * P
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    Thermo Fisher human hct 116 cells
    MVBs in T cells translocate to the IS. ( a ) J77 T cells were conjugated with SEE-pulsed or non-pulsed Raji B cells loaded with CMAC (blue). After 30 min, cells were fixed and stained for the MVB marker Hrs (red), CD3 and actin (both green). Plates show maximal projections of confocal images and the DIC images. ( b ) <t>CH7C17</t> T cells were conjugated with HA-loaded or non-loaded HOM2 B cells (blue). After 30 min, cells were fixed and stained for CD63 (green), CD3 and actin (both red). Plates show maximal projections of confocal images and the DIC images, scale bars applies to a and b . ( c ) Percentage of T and APC cells in which CD63, Hrs and CD3 relocalized to the T cell–APC contact area in the presence of HA peptide (filled bars) or its absence (open bars). Data are the arithmetic means±s.e.m. of four experiments. * P
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    Thermo Fisher moloney murine leukemia virus m mlv reverse transcriptase
    MVBs in T cells translocate to the IS. ( a ) J77 T cells were conjugated with SEE-pulsed or non-pulsed Raji B cells loaded with CMAC (blue). After 30 min, cells were fixed and stained for the MVB marker Hrs (red), CD3 and actin (both green). Plates show maximal projections of confocal images and the DIC images. ( b ) <t>CH7C17</t> T cells were conjugated with HA-loaded or non-loaded HOM2 B cells (blue). After 30 min, cells were fixed and stained for CD63 (green), CD3 and actin (both red). Plates show maximal projections of confocal images and the DIC images, scale bars applies to a and b . ( c ) Percentage of T and APC cells in which CD63, Hrs and CD3 relocalized to the T cell–APC contact area in the presence of HA peptide (filled bars) or its absence (open bars). Data are the arithmetic means±s.e.m. of four experiments. * P
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    MVBs in T cells translocate to the IS. ( a ) J77 T cells were conjugated with SEE-pulsed or non-pulsed Raji B cells loaded with CMAC (blue). After 30 min, cells were fixed and stained for the MVB marker Hrs (red), CD3 and actin (both green). Plates show maximal projections of confocal images and the DIC images. ( b ) <t>CH7C17</t> T cells were conjugated with HA-loaded or non-loaded HOM2 B cells (blue). After 30 min, cells were fixed and stained for CD63 (green), CD3 and actin (both red). Plates show maximal projections of confocal images and the DIC images, scale bars applies to a and b . ( c ) Percentage of T and APC cells in which CD63, Hrs and CD3 relocalized to the T cell–APC contact area in the presence of HA peptide (filled bars) or its absence (open bars). Data are the arithmetic means±s.e.m. of four experiments. * P
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    Illumina Inc genome analyser 2 platform
    MVBs in T cells translocate to the IS. ( a ) J77 T cells were conjugated with SEE-pulsed or non-pulsed Raji B cells loaded with CMAC (blue). After 30 min, cells were fixed and stained for the MVB marker Hrs (red), CD3 and actin (both green). Plates show maximal projections of confocal images and the DIC images. ( b ) <t>CH7C17</t> T cells were conjugated with HA-loaded or non-loaded HOM2 B cells (blue). After 30 min, cells were fixed and stained for CD63 (green), CD3 and actin (both red). Plates show maximal projections of confocal images and the DIC images, scale bars applies to a and b . ( c ) Percentage of T and APC cells in which CD63, Hrs and CD3 relocalized to the T cell–APC contact area in the presence of HA peptide (filled bars) or its absence (open bars). Data are the arithmetic means±s.e.m. of four experiments. * P
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    Thermo Fisher cdna ends
    MVBs in T cells translocate to the IS. ( a ) J77 T cells were conjugated with SEE-pulsed or non-pulsed Raji B cells loaded with CMAC (blue). After 30 min, cells were fixed and stained for the MVB marker Hrs (red), CD3 and actin (both green). Plates show maximal projections of confocal images and the DIC images. ( b ) <t>CH7C17</t> T cells were conjugated with HA-loaded or non-loaded HOM2 B cells (blue). After 30 min, cells were fixed and stained for CD63 (green), CD3 and actin (both red). Plates show maximal projections of confocal images and the DIC images, scale bars applies to a and b . ( c ) Percentage of T and APC cells in which CD63, Hrs and CD3 relocalized to the T cell–APC contact area in the presence of HA peptide (filled bars) or its absence (open bars). Data are the arithmetic means±s.e.m. of four experiments. * P
    Cdna Ends, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 9717 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Total RNA sample data from a lymphoblastoid cell-line. All data are normalized against let-7a data. Relative signal intensity of the let-7 family conventional ( a ) and ProDeG ( b ) probes are shown after Cy-3 labeled total RNA from a lymphoblastoid cell-line was hybridized at 34°C. The relative amount of each let-7 family miRNA was quantified using TaqMan® qRT-PCR assay ( c ).

    Journal: Nucleic Acids Research

    Article Title: Discriminating single-base difference miRNA expressions using microarray Probe Design Guru (ProDeG)

    doi: 10.1093/nar/gkm1165

    Figure Lengend Snippet: Total RNA sample data from a lymphoblastoid cell-line. All data are normalized against let-7a data. Relative signal intensity of the let-7 family conventional ( a ) and ProDeG ( b ) probes are shown after Cy-3 labeled total RNA from a lymphoblastoid cell-line was hybridized at 34°C. The relative amount of each let-7 family miRNA was quantified using TaqMan® qRT-PCR assay ( c ).

    Article Snippet: Hybridization experiments using total RNA of lymphoblastoid cell lines Total RNA from each human lymphoblastoid cell line was isolated with Trizol reagent (Invitrogen) according to the manufacturer's protocol (Invitrogen Cat No. 15596).

    Techniques: Labeling, Quantitative RT-PCR

    Cellular KAP1 regulates EBV lytic cycle in Burkitt Lymphoma cells. A-C. HH514-16 Burkitt lymphoma (BL) cells were transfected with scrambled siRNA (non-targeting control; open bars in B, C) or siRNA to KAP1 (black bars in B, C); D-F. BL cells were transfected with empty vector (open bars in E, F) or pFLAG-CMV2-KAP1 (black bars in E, F). Transfected cells were treated with NaB 24 hours after transfection, and harvested at indicated times (A, D), 24 hours (B, E), or 48 hours (C, F) post-treatment to determine relative amounts of ZEBRA, KAP1 and β-actin by immunoblotting (A, D), relative levels of transcripts from EBV lytic genes BZLF1 , BMRF1 and BFRF3 by qRT-PCR after normalization to 18S rRNA using the ΔΔC T method (B, E), and relative levels of cell-associated EBV DNA by qPCR (C, F). Error bars: SEM of 3 experiments with 4 technical replicates. Western blots are representative of 2 experiments. Numbers below blots indicate relative amounts of protein after normalization to β-actin.

    Journal: PLoS Pathogens

    Article Title: Chloroquine triggers Epstein-Barr virus replication through phosphorylation of KAP1/TRIM28 in Burkitt lymphoma cells

    doi: 10.1371/journal.ppat.1006249

    Figure Lengend Snippet: Cellular KAP1 regulates EBV lytic cycle in Burkitt Lymphoma cells. A-C. HH514-16 Burkitt lymphoma (BL) cells were transfected with scrambled siRNA (non-targeting control; open bars in B, C) or siRNA to KAP1 (black bars in B, C); D-F. BL cells were transfected with empty vector (open bars in E, F) or pFLAG-CMV2-KAP1 (black bars in E, F). Transfected cells were treated with NaB 24 hours after transfection, and harvested at indicated times (A, D), 24 hours (B, E), or 48 hours (C, F) post-treatment to determine relative amounts of ZEBRA, KAP1 and β-actin by immunoblotting (A, D), relative levels of transcripts from EBV lytic genes BZLF1 , BMRF1 and BFRF3 by qRT-PCR after normalization to 18S rRNA using the ΔΔC T method (B, E), and relative levels of cell-associated EBV DNA by qPCR (C, F). Error bars: SEM of 3 experiments with 4 technical replicates. Western blots are representative of 2 experiments. Numbers below blots indicate relative amounts of protein after normalization to β-actin.

    Article Snippet: BACmid p2089 was a gift from Professor Henri-Jacques Delecluse [ ]. siRNAs targeting human KAP1 and ATM transcripts and scrambled/non-targeting siRNA were purchased from Santa Cruz Biotechnology (sc-38550, sc-29761 and sc-37007) and Dharmacon (D-001206-13-05 and M-005046-01) and reconstituted with nuclease free water.

    Techniques: Transfection, Plasmid Preparation, Quantitative RT-PCR, Real-time Polymerase Chain Reaction, Western Blot

    ATM induces phosphorylation of KAP1 at S824 upon exposure to lytic trigger. A. HH514-16 BL cells were treated with increasing amounts of PI3 kinase-related kinase inhibitors (KU-55933 or Torin1; left panels) or PI3KK inhibitors plus NaB (right panels). After 24 hours, cells were harvested and lysates analyzed via immunoblotting with the antibodies indicated. B. HH514-16 cells were treated with NaB (upper panel) or NaB plus KU-55933 (1μM; middle panel) or NaB plus Torin1 (0.5μM; lower panel) for 24 hours and stained with anti-phospho KAP1 (S824) plus anti-ZEBRA antibodies and visualized at 1000X magnification. C. HH514-16 cells were transfected with scrambled siRNA or siRNA to ATM , treated with NaB after 24 hours and harvested after another 24 hours. Cell lysates were analyzed by immunoblotting with the antibodies indicated. Numbers below p-S824 KAP1 blot indicate relative amounts of pKAP1 after normalization to total KAP1. D. HH514-16-derived CLIX-FZ cells were exposed to doxycycline (upper panel) or doxycycline plus KU-55933 (lower panel) for 24 hours and stained with anti-p-S824 KAP1 plus anti-ZEBRA antibodies and visualized at 1000X magnification. E. Lymphoblastoid cells were transfected with pHD1013-Z plasmid and simultaneously treated with vehicle (-KU) or KU-55933. Treated cells were harvested 24 hours later and subjected to staining with anti-phospho KAP1 (S824) plus anti-ZEBRA antibodies and visualized at 1000X magnification. Experiments were performed twice.

    Journal: PLoS Pathogens

    Article Title: Chloroquine triggers Epstein-Barr virus replication through phosphorylation of KAP1/TRIM28 in Burkitt lymphoma cells

    doi: 10.1371/journal.ppat.1006249

    Figure Lengend Snippet: ATM induces phosphorylation of KAP1 at S824 upon exposure to lytic trigger. A. HH514-16 BL cells were treated with increasing amounts of PI3 kinase-related kinase inhibitors (KU-55933 or Torin1; left panels) or PI3KK inhibitors plus NaB (right panels). After 24 hours, cells were harvested and lysates analyzed via immunoblotting with the antibodies indicated. B. HH514-16 cells were treated with NaB (upper panel) or NaB plus KU-55933 (1μM; middle panel) or NaB plus Torin1 (0.5μM; lower panel) for 24 hours and stained with anti-phospho KAP1 (S824) plus anti-ZEBRA antibodies and visualized at 1000X magnification. C. HH514-16 cells were transfected with scrambled siRNA or siRNA to ATM , treated with NaB after 24 hours and harvested after another 24 hours. Cell lysates were analyzed by immunoblotting with the antibodies indicated. Numbers below p-S824 KAP1 blot indicate relative amounts of pKAP1 after normalization to total KAP1. D. HH514-16-derived CLIX-FZ cells were exposed to doxycycline (upper panel) or doxycycline plus KU-55933 (lower panel) for 24 hours and stained with anti-p-S824 KAP1 plus anti-ZEBRA antibodies and visualized at 1000X magnification. E. Lymphoblastoid cells were transfected with pHD1013-Z plasmid and simultaneously treated with vehicle (-KU) or KU-55933. Treated cells were harvested 24 hours later and subjected to staining with anti-phospho KAP1 (S824) plus anti-ZEBRA antibodies and visualized at 1000X magnification. Experiments were performed twice.

    Article Snippet: BACmid p2089 was a gift from Professor Henri-Jacques Delecluse [ ]. siRNAs targeting human KAP1 and ATM transcripts and scrambled/non-targeting siRNA were purchased from Santa Cruz Biotechnology (sc-38550, sc-29761 and sc-37007) and Dharmacon (D-001206-13-05 and M-005046-01) and reconstituted with nuclease free water.

    Techniques: Staining, Transfection, Derivative Assay, Plasmid Preparation

    MVBs in T cells translocate to the IS. ( a ) J77 T cells were conjugated with SEE-pulsed or non-pulsed Raji B cells loaded with CMAC (blue). After 30 min, cells were fixed and stained for the MVB marker Hrs (red), CD3 and actin (both green). Plates show maximal projections of confocal images and the DIC images. ( b ) CH7C17 T cells were conjugated with HA-loaded or non-loaded HOM2 B cells (blue). After 30 min, cells were fixed and stained for CD63 (green), CD3 and actin (both red). Plates show maximal projections of confocal images and the DIC images, scale bars applies to a and b . ( c ) Percentage of T and APC cells in which CD63, Hrs and CD3 relocalized to the T cell–APC contact area in the presence of HA peptide (filled bars) or its absence (open bars). Data are the arithmetic means±s.e.m. of four experiments. * P

    Journal: Nature Communications

    Article Title: Unidirectional transfer of microRNA-loaded exosomes from T cells to antigen-presenting cells

    doi: 10.1038/ncomms1285

    Figure Lengend Snippet: MVBs in T cells translocate to the IS. ( a ) J77 T cells were conjugated with SEE-pulsed or non-pulsed Raji B cells loaded with CMAC (blue). After 30 min, cells were fixed and stained for the MVB marker Hrs (red), CD3 and actin (both green). Plates show maximal projections of confocal images and the DIC images. ( b ) CH7C17 T cells were conjugated with HA-loaded or non-loaded HOM2 B cells (blue). After 30 min, cells were fixed and stained for CD63 (green), CD3 and actin (both red). Plates show maximal projections of confocal images and the DIC images, scale bars applies to a and b . ( c ) Percentage of T and APC cells in which CD63, Hrs and CD3 relocalized to the T cell–APC contact area in the presence of HA peptide (filled bars) or its absence (open bars). Data are the arithmetic means±s.e.m. of four experiments. * P

    Article Snippet: 2 Vβ8) and CH7C17 (Vβ3 TCR specific for HA peptide) and the lymphoblastoid B-cell lines Raji (Burkitt lymphoma), HOM-2 (HLA-DR1 EBV-transformed) and BLS-1 (HLA class II-null B-LCL, generated from cells of patients with type II Bare lymphocyte syndrome) were cultured in RPMI (Sigma) containing 10% fetal bovine serum (Invitrogen).

    Techniques: Staining, Marker

    Exosomal miRNA-335 is transferred from T cell to APC in an Ag-specific manner. ( a ) Levels of miR-335 were assessed by quantitative reverse transcription PCR (qRT–PCR) in primary dendritic cells and T lymphoblasts, and in Raji and J77 cells. J77-CD63-GFP cells were stably transduced with miR-335 (J-335 cells) or miR-101 (J-101 cells), and miR-335 levels were determined by qRT–PCR in cells and derived exosomes. Data are representative of three experiments (mean and s.e.m.) ( b ) miR-335 levels in SEE-primed Raji cells sorted 24 h after conjugation with J-335 cells. J-101 were used as control donor cells. Left panel, Data are representative of seven independent experiments (mean and s.e.m), P =0.014 (one-sample t -test). Right panel, n =5 independent experiments; P =0.04 (one-sample t -test); error bars represent s.e.m. ( c ) miR-335 levels in HA-primed HOM-2 cells sorted 24 h after conjugation with CH7C17 cells overexpressing miR-335 (C-335). CH7C17 cells overexpressing miR-101 (C-101) were used as control donor cells. Data are representative of five experiments (mean and s.e.m.). ( d ) miR-335 and miR-92a levels in SEE-primed Raji cells sorted 24 h after conjugation with primary T lymphoblasts-expressing miR-335 and miR-92a endogenously. Data are representative of three experiments (mean and s.e.m.), * P =0.026 (one-sample t -test) AU, arbitrary units.

    Journal: Nature Communications

    Article Title: Unidirectional transfer of microRNA-loaded exosomes from T cells to antigen-presenting cells

    doi: 10.1038/ncomms1285

    Figure Lengend Snippet: Exosomal miRNA-335 is transferred from T cell to APC in an Ag-specific manner. ( a ) Levels of miR-335 were assessed by quantitative reverse transcription PCR (qRT–PCR) in primary dendritic cells and T lymphoblasts, and in Raji and J77 cells. J77-CD63-GFP cells were stably transduced with miR-335 (J-335 cells) or miR-101 (J-101 cells), and miR-335 levels were determined by qRT–PCR in cells and derived exosomes. Data are representative of three experiments (mean and s.e.m.) ( b ) miR-335 levels in SEE-primed Raji cells sorted 24 h after conjugation with J-335 cells. J-101 were used as control donor cells. Left panel, Data are representative of seven independent experiments (mean and s.e.m), P =0.014 (one-sample t -test). Right panel, n =5 independent experiments; P =0.04 (one-sample t -test); error bars represent s.e.m. ( c ) miR-335 levels in HA-primed HOM-2 cells sorted 24 h after conjugation with CH7C17 cells overexpressing miR-335 (C-335). CH7C17 cells overexpressing miR-101 (C-101) were used as control donor cells. Data are representative of five experiments (mean and s.e.m.). ( d ) miR-335 and miR-92a levels in SEE-primed Raji cells sorted 24 h after conjugation with primary T lymphoblasts-expressing miR-335 and miR-92a endogenously. Data are representative of three experiments (mean and s.e.m.), * P =0.026 (one-sample t -test) AU, arbitrary units.

    Article Snippet: 2 Vβ8) and CH7C17 (Vβ3 TCR specific for HA peptide) and the lymphoblastoid B-cell lines Raji (Burkitt lymphoma), HOM-2 (HLA-DR1 EBV-transformed) and BLS-1 (HLA class II-null B-LCL, generated from cells of patients with type II Bare lymphocyte syndrome) were cultured in RPMI (Sigma) containing 10% fetal bovine serum (Invitrogen).

    Techniques: Polymerase Chain Reaction, Quantitative RT-PCR, Stable Transfection, Transduction, Derivative Assay, Conjugation Assay, Expressing