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  • 99
    ATCC cell lines e1a e1b trans complementing hek 293 cell line
    Cell Lines E1a E1b Trans Complementing Hek 293 Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Thermo Fisher bcl2 adenovirus e1b 19kda interacting protein 3
    Bcl2 Adenovirus E1b 19kda Interacting Protein 3, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene minimal e1b promoter luciferase gene
    Minimal E1b Promoter Luciferase Gene, supplied by Stratagene, used in various techniques. Bioz Stars score: 88/100, based on 59 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Genewiz e1b gene
    Viral DNA synthesis is inhibited in AdEasyE1Sub19-infected cells treated with IFN. (A) HFFs were infected with 30 PFU/cell AdEasyE1 (wild type [WT]), AdEasyE1Δ2347 (ΔE1B), or AdEasyE1Sub19 (S19) for the periods indicated or mock infected (M). The proteins listed at the right were examined by immunoblotting of total cell lysates as described in Materials and Methods. (B) HFFs were infected with 30 PFU/cell AdEasyE1 (wild type [WT]) or AdEasyE1Sub19 (S19) for 36 h or mock infected (mock), and the <t>E1B</t> 55-kDa protein was visualized by immunofluorescence as described in Materials and Methods. DAPI, 4′,6-diamidino-2-phenylindole. (C) HFFs were treated with IFN or untreated (BSA) and infected with 3 PFU/cell of the virus indicated for 2 or 72 h. Viral DNA concentrations were measured by quantitative PCR, with cellular β-actin DNA used as an internal control, as described in Materials and Methods, and are expressed relative to the input (2 h) concentrations. The values represent the averages of two independent experiments, with cumulative standard deviations indicated by the error bars.
    E1b Gene, supplied by Genewiz, used in various techniques. Bioz Stars score: 85/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    ZIRC Inc uas e1b
    Phenotypic characterization of a stable transgenic line. ( a ) Graphic representation of the DNA constructs used for transgenesis and breeding. ( b ) A 24 h.p.f. embryo expresses Smoa1-EGFP in the forebrain and retinal neuroblasts. A 96 h.p.f. double transgenic embryo derived from crossing of the Tg(krt5:Gal4VP16;14 × <t>UAS:smoa1-EGFP)</t> and the <t>Tg(UAS-E1b:nfsB-mCherry)</t> c264 stable lines expressed GFP ( c ) and mCherry ( d ) in the ONs (arrows). ( e ) A 6-month-old wild-type and ( f ) a transgenic fish with a gross tumor in its left eye (arrow). ( g ) Wild-type fish formed a normal optic chiasm, whereas ( h ) transgenic fish failed to form optic chiasm and showed dramatic expansion of the ONH region (arrow). ( i ) Dissected brain and eyes showing an enlarged ON associating with a gross eye tumor (arrow).
    Uas E1b, supplied by ZIRC Inc, used in various techniques. Bioz Stars score: 85/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc bcl2 adenovirus e1b 19
    Phenotypic characterization of a stable transgenic line. ( a ) Graphic representation of the DNA constructs used for transgenesis and breeding. ( b ) A 24 h.p.f. embryo expresses Smoa1-EGFP in the forebrain and retinal neuroblasts. A 96 h.p.f. double transgenic embryo derived from crossing of the Tg(krt5:Gal4VP16;14 × <t>UAS:smoa1-EGFP)</t> and the <t>Tg(UAS-E1b:nfsB-mCherry)</t> c264 stable lines expressed GFP ( c ) and mCherry ( d ) in the ONs (arrows). ( e ) A 6-month-old wild-type and ( f ) a transgenic fish with a gross tumor in its left eye (arrow). ( g ) Wild-type fish formed a normal optic chiasm, whereas ( h ) transgenic fish failed to form optic chiasm and showed dramatic expansion of the ONH region (arrow). ( i ) Dissected brain and eyes showing an enlarged ON associating with a gross eye tumor (arrow).
    Bcl2 Adenovirus E1b 19, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 98/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Bethyl rabbit anti e1b ap5 antibody affinity purified
    Phenotypic characterization of a stable transgenic line. ( a ) Graphic representation of the DNA constructs used for transgenesis and breeding. ( b ) A 24 h.p.f. embryo expresses Smoa1-EGFP in the forebrain and retinal neuroblasts. A 96 h.p.f. double transgenic embryo derived from crossing of the Tg(krt5:Gal4VP16;14 × <t>UAS:smoa1-EGFP)</t> and the <t>Tg(UAS-E1b:nfsB-mCherry)</t> c264 stable lines expressed GFP ( c ) and mCherry ( d ) in the ONs (arrows). ( e ) A 6-month-old wild-type and ( f ) a transgenic fish with a gross tumor in its left eye (arrow). ( g ) Wild-type fish formed a normal optic chiasm, whereas ( h ) transgenic fish failed to form optic chiasm and showed dramatic expansion of the ONH region (arrow). ( i ) Dissected brain and eyes showing an enlarged ON associating with a gross eye tumor (arrow).
    Rabbit Anti E1b Ap5 Antibody Affinity Purified, supplied by Bethyl, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Onyx Pharmaceuticals e1b 55 kda gene deleted ad5 mutant dl1520
    Phenotypic characterization of a stable transgenic line. ( a ) Graphic representation of the DNA constructs used for transgenesis and breeding. ( b ) A 24 h.p.f. embryo expresses Smoa1-EGFP in the forebrain and retinal neuroblasts. A 96 h.p.f. double transgenic embryo derived from crossing of the Tg(krt5:Gal4VP16;14 × <t>UAS:smoa1-EGFP)</t> and the <t>Tg(UAS-E1b:nfsB-mCherry)</t> c264 stable lines expressed GFP ( c ) and mCherry ( d ) in the ONs (arrows). ( e ) A 6-month-old wild-type and ( f ) a transgenic fish with a gross tumor in its left eye (arrow). ( g ) Wild-type fish formed a normal optic chiasm, whereas ( h ) transgenic fish failed to form optic chiasm and showed dramatic expansion of the ONH region (arrow). ( i ) Dissected brain and eyes showing an enlarged ON associating with a gross eye tumor (arrow).
    E1b 55 Kda Gene Deleted Ad5 Mutant Dl1520, supplied by Onyx Pharmaceuticals, used in various techniques. Bioz Stars score: 88/100, based on 63 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti bcl2 adenovirus e1b 19 kda interacting protein 3
    Phenotypic characterization of a stable transgenic line. ( a ) Graphic representation of the DNA constructs used for transgenesis and breeding. ( b ) A 24 h.p.f. embryo expresses Smoa1-EGFP in the forebrain and retinal neuroblasts. A 96 h.p.f. double transgenic embryo derived from crossing of the Tg(krt5:Gal4VP16;14 × <t>UAS:smoa1-EGFP)</t> and the <t>Tg(UAS-E1b:nfsB-mCherry)</t> c264 stable lines expressed GFP ( c ) and mCherry ( d ) in the ONs (arrows). ( e ) A 6-month-old wild-type and ( f ) a transgenic fish with a gross tumor in its left eye (arrow). ( g ) Wild-type fish formed a normal optic chiasm, whereas ( h ) transgenic fish failed to form optic chiasm and showed dramatic expansion of the ONH region (arrow). ( i ) Dissected brain and eyes showing an enlarged ON associating with a gross eye tumor (arrow).
    Anti Bcl2 Adenovirus E1b 19 Kda Interacting Protein 3, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Calbiotech anti adenovirus type 5 e1b 55k ab 1 rat monoclonal antibody
    Phenotypic characterization of a stable transgenic line. ( a ) Graphic representation of the DNA constructs used for transgenesis and breeding. ( b ) A 24 h.p.f. embryo expresses Smoa1-EGFP in the forebrain and retinal neuroblasts. A 96 h.p.f. double transgenic embryo derived from crossing of the Tg(krt5:Gal4VP16;14 × <t>UAS:smoa1-EGFP)</t> and the <t>Tg(UAS-E1b:nfsB-mCherry)</t> c264 stable lines expressed GFP ( c ) and mCherry ( d ) in the ONs (arrows). ( e ) A 6-month-old wild-type and ( f ) a transgenic fish with a gross tumor in its left eye (arrow). ( g ) Wild-type fish formed a normal optic chiasm, whereas ( h ) transgenic fish failed to form optic chiasm and showed dramatic expansion of the ONH region (arrow). ( i ) Dissected brain and eyes showing an enlarged ON associating with a gross eye tumor (arrow).
    Anti Adenovirus Type 5 E1b 55k Ab 1 Rat Monoclonal Antibody, supplied by Calbiotech, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti e1b ap5 antibody
    KSHV ORF57 interacts with PABPC1. (A) ORF57 interacts with PABPC1, but not with <t>E1B-AP5,</t> as shown by coimmunoprecipitation assays. Total cell extracts from TREx BCBL1-Rta cells induced by Dox (1 μg/ml) and treated with a mixture of RNase A and T1 were immunoprecipitated with preimmune mouse IgG- or monoclonal anti-ORF57 antibody-coated beads and examined for PABPC1 and E1B-AP5 by Western blotting (left). Conversely, the same extracts treated with a mixture of RNase A and T1 were immunoprecipitated with nonspecific rabbit IgG- or polyclonal anti-PABPC1 or anti-E1B-AP5 antibody-coated beads and blotted for ORF57 by Western blotting (right). (B) PABPC1 directly interacts with ORF57, as shown by GST pulldown assays. Full-length PABPC1 protein with an N-terminal GST tag or untagged GST was immobilized on glutathione-Sepharose 4B beads and incubated at 4°C overnight in IP buffer complemented with KSHV ORF57 protein containing a C-terminal FLAG tag that was expressed from baculovirus. The glutathione-Sepharose 4B beads without any GST protein were used as an additional negative control. After binding, the beads were washed five times with IP buffer, resuspended in 2× SDS sample buffer, and analyzed by Western blotting with two antibodies, an anti-GST antibody recognizing PABPC1 and an anti-FLAG antibody that recognizes the epitope-tagged ORF57.
    Anti E1b Ap5 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam bcl 2 adenovirus e1b 19 kda interacting protein 3 like bnip3
    KSHV ORF57 interacts with PABPC1. (A) ORF57 interacts with PABPC1, but not with <t>E1B-AP5,</t> as shown by coimmunoprecipitation assays. Total cell extracts from TREx BCBL1-Rta cells induced by Dox (1 μg/ml) and treated with a mixture of RNase A and T1 were immunoprecipitated with preimmune mouse IgG- or monoclonal anti-ORF57 antibody-coated beads and examined for PABPC1 and E1B-AP5 by Western blotting (left). Conversely, the same extracts treated with a mixture of RNase A and T1 were immunoprecipitated with nonspecific rabbit IgG- or polyclonal anti-PABPC1 or anti-E1B-AP5 antibody-coated beads and blotted for ORF57 by Western blotting (right). (B) PABPC1 directly interacts with ORF57, as shown by GST pulldown assays. Full-length PABPC1 protein with an N-terminal GST tag or untagged GST was immobilized on glutathione-Sepharose 4B beads and incubated at 4°C overnight in IP buffer complemented with KSHV ORF57 protein containing a C-terminal FLAG tag that was expressed from baculovirus. The glutathione-Sepharose 4B beads without any GST protein were used as an additional negative control. After binding, the beads were washed five times with IP buffer, resuspended in 2× SDS sample buffer, and analyzed by Western blotting with two antibodies, an anti-GST antibody recognizing PABPC1 and an anti-FLAG antibody that recognizes the epitope-tagged ORF57.
    Bcl 2 Adenovirus E1b 19 Kda Interacting Protein 3 Like Bnip3, supplied by Abcam, used in various techniques. Bioz Stars score: 96/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc bcl 2 adenovirus e1b 19 k
    KSHV ORF57 interacts with PABPC1. (A) ORF57 interacts with PABPC1, but not with <t>E1B-AP5,</t> as shown by coimmunoprecipitation assays. Total cell extracts from TREx BCBL1-Rta cells induced by Dox (1 μg/ml) and treated with a mixture of RNase A and T1 were immunoprecipitated with preimmune mouse IgG- or monoclonal anti-ORF57 antibody-coated beads and examined for PABPC1 and E1B-AP5 by Western blotting (left). Conversely, the same extracts treated with a mixture of RNase A and T1 were immunoprecipitated with nonspecific rabbit IgG- or polyclonal anti-PABPC1 or anti-E1B-AP5 antibody-coated beads and blotted for ORF57 by Western blotting (right). (B) PABPC1 directly interacts with ORF57, as shown by GST pulldown assays. Full-length PABPC1 protein with an N-terminal GST tag or untagged GST was immobilized on glutathione-Sepharose 4B beads and incubated at 4°C overnight in IP buffer complemented with KSHV ORF57 protein containing a C-terminal FLAG tag that was expressed from baculovirus. The glutathione-Sepharose 4B beads without any GST protein were used as an additional negative control. After binding, the beads were washed five times with IP buffer, resuspended in 2× SDS sample buffer, and analyzed by Western blotting with two antibodies, an anti-GST antibody recognizing PABPC1 and an anti-FLAG antibody that recognizes the epitope-tagged ORF57.
    Bcl 2 Adenovirus E1b 19 K, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher rabbit serum e1b
    KSHV ORF57 interacts with PABPC1. (A) ORF57 interacts with PABPC1, but not with <t>E1B-AP5,</t> as shown by coimmunoprecipitation assays. Total cell extracts from TREx BCBL1-Rta cells induced by Dox (1 μg/ml) and treated with a mixture of RNase A and T1 were immunoprecipitated with preimmune mouse IgG- or monoclonal anti-ORF57 antibody-coated beads and examined for PABPC1 and E1B-AP5 by Western blotting (left). Conversely, the same extracts treated with a mixture of RNase A and T1 were immunoprecipitated with nonspecific rabbit IgG- or polyclonal anti-PABPC1 or anti-E1B-AP5 antibody-coated beads and blotted for ORF57 by Western blotting (right). (B) PABPC1 directly interacts with ORF57, as shown by GST pulldown assays. Full-length PABPC1 protein with an N-terminal GST tag or untagged GST was immobilized on glutathione-Sepharose 4B beads and incubated at 4°C overnight in IP buffer complemented with KSHV ORF57 protein containing a C-terminal FLAG tag that was expressed from baculovirus. The glutathione-Sepharose 4B beads without any GST protein were used as an additional negative control. After binding, the beads were washed five times with IP buffer, resuspended in 2× SDS sample buffer, and analyzed by Western blotting with two antibodies, an anti-GST antibody recognizing PABPC1 and an anti-FLAG antibody that recognizes the epitope-tagged ORF57.
    Rabbit Serum E1b, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Oncogene Science Inc e1b 19 kda
    HCMV IE1 and IE2 genes and their products are not present in transformed BRK cell lines. ( A ) Lack of expression of the IE1 or IE2 protein in transformed BRK cell lines when assayed by immunoprecipitation of 35 S-labeled cell extracts using monoclonal antibody mAB810. A-1 is a cell line established by transfection with the E1A-expressing plasmid alone. A/1/2–1 to A/1/2–10 are cell lines transformed by E1A plus IE1 and IE2. Lanes designated HCMV INF display immunoprecipitates from HCMV-infected human foreskin fibroblasts; lane 13 is an exposure 1/8 as long as that presented for lanes 1–12. Numbers to the left of the gel mark the positions of marker proteins whose sizes are indicated in <t>kDa.</t> ( B ) Southern blot analysis showing the lack of IE1 - or IE2 -specific DNA in E1A/IE1/IE2-induced transformants (lanes 1–10). A/B-1 and A/B-2 are cell lines transformed with E1A and <t>E1B-19-kDa.</t> HeLa, 293, and HP1.14 (a derivative HeLa cells expressing IE1) are included as controls. pCGN-IE1 and pCGN-IE2 are IE1 and IE2 expression plasmids, respectively, in amounts equivalent to the rest of the samples, assuming they contain one copy of each of the HCMV genes per cell. Numbers to the left of the gel are marker DNA sizes in kb. ( C ) Expression of E1A protein as determined by immunoprecipitation followed by immunoblot analysis, both with monoclonal antibody M73 to E1A. Lanes are labeled as in A and B . Numbers to the left of the gel are marker protein sizes in kDa. Autoradiograms were scanned and cropped using Photoshop and figures were prepared using Freehand software.
    E1b 19 Kda, supplied by Oncogene Science Inc, used in various techniques. Bioz Stars score: 80/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Oncogene Science Inc e1b 19k rat mab
    HCMV IE1 and IE2 genes and their products are not present in transformed BRK cell lines. ( A ) Lack of expression of the IE1 or IE2 protein in transformed BRK cell lines when assayed by immunoprecipitation of 35 S-labeled cell extracts using monoclonal antibody mAB810. A-1 is a cell line established by transfection with the E1A-expressing plasmid alone. A/1/2–1 to A/1/2–10 are cell lines transformed by E1A plus IE1 and IE2. Lanes designated HCMV INF display immunoprecipitates from HCMV-infected human foreskin fibroblasts; lane 13 is an exposure 1/8 as long as that presented for lanes 1–12. Numbers to the left of the gel mark the positions of marker proteins whose sizes are indicated in <t>kDa.</t> ( B ) Southern blot analysis showing the lack of IE1 - or IE2 -specific DNA in E1A/IE1/IE2-induced transformants (lanes 1–10). A/B-1 and A/B-2 are cell lines transformed with E1A and <t>E1B-19-kDa.</t> HeLa, 293, and HP1.14 (a derivative HeLa cells expressing IE1) are included as controls. pCGN-IE1 and pCGN-IE2 are IE1 and IE2 expression plasmids, respectively, in amounts equivalent to the rest of the samples, assuming they contain one copy of each of the HCMV genes per cell. Numbers to the left of the gel are marker DNA sizes in kb. ( C ) Expression of E1A protein as determined by immunoprecipitation followed by immunoblot analysis, both with monoclonal antibody M73 to E1A. Lanes are labeled as in A and B . Numbers to the left of the gel are marker protein sizes in kDa. Autoradiograms were scanned and cropped using Photoshop and figures were prepared using Freehand software.
    E1b 19k Rat Mab, supplied by Oncogene Science Inc, 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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    becton dickinson e1b 19k
    HCMV IE1 and IE2 genes and their products are not present in transformed BRK cell lines. ( A ) Lack of expression of the IE1 or IE2 protein in transformed BRK cell lines when assayed by immunoprecipitation of 35 S-labeled cell extracts using monoclonal antibody mAB810. A-1 is a cell line established by transfection with the E1A-expressing plasmid alone. A/1/2–1 to A/1/2–10 are cell lines transformed by E1A plus IE1 and IE2. Lanes designated HCMV INF display immunoprecipitates from HCMV-infected human foreskin fibroblasts; lane 13 is an exposure 1/8 as long as that presented for lanes 1–12. Numbers to the left of the gel mark the positions of marker proteins whose sizes are indicated in <t>kDa.</t> ( B ) Southern blot analysis showing the lack of IE1 - or IE2 -specific DNA in E1A/IE1/IE2-induced transformants (lanes 1–10). A/B-1 and A/B-2 are cell lines transformed with E1A and <t>E1B-19-kDa.</t> HeLa, 293, and HP1.14 (a derivative HeLa cells expressing IE1) are included as controls. pCGN-IE1 and pCGN-IE2 are IE1 and IE2 expression plasmids, respectively, in amounts equivalent to the rest of the samples, assuming they contain one copy of each of the HCMV genes per cell. Numbers to the left of the gel are marker DNA sizes in kb. ( C ) Expression of E1A protein as determined by immunoprecipitation followed by immunoblot analysis, both with monoclonal antibody M73 to E1A. Lanes are labeled as in A and B . Numbers to the left of the gel are marker protein sizes in kDa. Autoradiograms were scanned and cropped using Photoshop and figures were prepared using Freehand software.
    E1b 19k, supplied by becton dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Genewiz e1b 55 kda insert
    Viral DNA synthesis is inhibited in AdEasyE1Sub19-infected cells treated with IFN. (A) HFFs were infected with 30 PFU/cell AdEasyE1 (wild type [WT]), AdEasyE1Δ2347 (ΔE1B), or AdEasyE1Sub19 (S19) for the periods indicated or mock infected (M). The proteins listed at the right were examined by immunoblotting of total cell lysates as described in Materials and Methods. (B) HFFs were infected with 30 PFU/cell AdEasyE1 (wild type [WT]) or AdEasyE1Sub19 (S19) for 36 h or mock infected (mock), and the E1B <t>55-kDa</t> protein was visualized by immunofluorescence as described in Materials and Methods. DAPI, 4′,6-diamidino-2-phenylindole. (C) HFFs were treated with IFN or untreated (BSA) and infected with 3 PFU/cell of the virus indicated for 2 or 72 h. Viral DNA concentrations were measured by quantitative PCR, with cellular β-actin DNA used as an internal control, as described in Materials and Methods, and are expressed relative to the input (2 h) concentrations. The values represent the averages of two independent experiments, with cumulative standard deviations indicated by the error bars.
    E1b 55 Kda Insert, supplied by Genewiz, used in various techniques. Bioz Stars score: 85/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    OriGene e1b 55 k ap 000199 expression vectors
    Viral DNA synthesis is inhibited in AdEasyE1Sub19-infected cells treated with IFN. (A) HFFs were infected with 30 PFU/cell AdEasyE1 (wild type [WT]), AdEasyE1Δ2347 (ΔE1B), or AdEasyE1Sub19 (S19) for the periods indicated or mock infected (M). The proteins listed at the right were examined by immunoblotting of total cell lysates as described in Materials and Methods. (B) HFFs were infected with 30 PFU/cell AdEasyE1 (wild type [WT]) or AdEasyE1Sub19 (S19) for 36 h or mock infected (mock), and the E1B <t>55-kDa</t> protein was visualized by immunofluorescence as described in Materials and Methods. DAPI, 4′,6-diamidino-2-phenylindole. (C) HFFs were treated with IFN or untreated (BSA) and infected with 3 PFU/cell of the virus indicated for 2 or 72 h. Viral DNA concentrations were measured by quantitative PCR, with cellular β-actin DNA used as an internal control, as described in Materials and Methods, and are expressed relative to the input (2 h) concentrations. The values represent the averages of two independent experiments, with cumulative standard deviations indicated by the error bars.
    E1b 55 K Ap 000199 Expression Vectors, supplied by OriGene, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Biomatik e1b 55k coding region
    E4-ORF3 induces SUMO conjugation to <t>E1B-55K,</t> Nbs1, and Mre11. (A) HeLa cells infected with dl 355 (ΔE4-ORF6) or dl 355- in ORF3 (ΔE4-ORF3/ΔE4-ORF6) were immunostained for E1B-55K and E2-DBP or E4-ORF3 at 12 hpi. (B) His 6 -tagged SUMO-1 or SUMO-3-expressing HeLa cells were infected with indicating amounts of dl 355 or dl 355- in ORF3. At 12 hpi, SUMO conjugates were captured by Ni-NTA agarose beads and analyzed by Western blotting with the anti-E1B-55K antibody. SUMO-conjugated RanGAP1 was shown as a control for SUMO capture. Protein levels of E1B-55K and E4-ORF3 from cell lysates were determined by Western blotting. (C) GST-tagged Ad5 E1B-55K was incubated with SUMO-1/E1/E2 and the indicated concentrations of Ad5 E4-ORF3. The levels of SUMO conjugation were analyzed by Western blotting with the anti-E1B-55K antibody. (D) GST-tagged Nbs1 and Mre11 were incubated with SUMO E1/E2 and SUMO-1 or SUMO-3 in the absence or presence of Ad5 E4-ORF3 (3 μM). The reaction mixture was analyzed by Western blotting with anti-Nbs1 and anti-Mre11 antibodies.
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    Oncogene Science Inc e1b 55k protein specific rat monoclonal antibody 9c10
    Fusion of human cells to mouse A9 cells expressing the <t>E1B-55K</t> and E4orf6 genes permits the E1B-55K protein to enter mouse A9 cell nuclei. Expression of the E1B-55K and E4orf6 proteins was established in mouse A9 cells with the recombinant vaccinia virus T7 infection-transfection system. HeLa cells were harvested by trypsin treatment and allowed to attach to the substrate with adherent A9 cells. The HeLa and A9 cells were fused 8 h after transfection with PEG. The resulting heterokaryons were maintained in the presence of cycloheximide (100 μg/ml) for 4 h before processing for double-label immunofluorescence. (A) Phase contrast image showing two heterokaryons; (B) DAPI-stained image showing DNA; (C) staining for the large subunit of the human Ku protein specifically labels the HeLa cell nuclei; (D) staining for the E1B-55K protein with the rat monoclonal antibody <t>9C10.</t> A9 cell nuclei are marked by arrowheads.
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    Parke-Davis e1b 55kd gene
    Fusion of human cells to mouse A9 cells expressing the <t>E1B-55K</t> and E4orf6 genes permits the E1B-55K protein to enter mouse A9 cell nuclei. Expression of the E1B-55K and E4orf6 proteins was established in mouse A9 cells with the recombinant vaccinia virus T7 infection-transfection system. HeLa cells were harvested by trypsin treatment and allowed to attach to the substrate with adherent A9 cells. The HeLa and A9 cells were fused 8 h after transfection with PEG. The resulting heterokaryons were maintained in the presence of cycloheximide (100 μg/ml) for 4 h before processing for double-label immunofluorescence. (A) Phase contrast image showing two heterokaryons; (B) DAPI-stained image showing DNA; (C) staining for the large subunit of the human Ku protein specifically labels the HeLa cell nuclei; (D) staining for the E1B-55K protein with the rat monoclonal antibody <t>9C10.</t> A9 cell nuclei are marked by arrowheads.
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    Thermo Fisher e1b ap5
    (A) Localization of <t>E1B-AP5,</t> ATRIP, and RPA32 in mock-infected interphase A549 cells. Cells were grown on glass coverslips and then treated with preextraction buffer and fixed with 4% (wt/vol) paraformaldehyde as described in Materials and Methods. Antigens were detected using the appropriate antibodies. (B) Localization of pTP, DBP, and RPA32 at viral replication centers in Ad5-infected A549 cells. Twenty-four hours postinfection with 10 PFU/cell wt Ad5, cells were treated with preextraction buffer and then fixed with 4% (wt/vol) paraformaldehyde, whereupon antigens were detected using the appropriate reagents. Colocalization images were recorded using a Zeiss LSM510-Meta laser scanning confocal microscope. Nuclei are stained with DAPI and are shown in blue.
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    Xenobiotics e1b expression modulators
    Effect of overexpression and siRNA knockdown of NRF2 on <t>E1b</t> expression
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    Image Search Results


    Viral DNA synthesis is inhibited in AdEasyE1Sub19-infected cells treated with IFN. (A) HFFs were infected with 30 PFU/cell AdEasyE1 (wild type [WT]), AdEasyE1Δ2347 (ΔE1B), or AdEasyE1Sub19 (S19) for the periods indicated or mock infected (M). The proteins listed at the right were examined by immunoblotting of total cell lysates as described in Materials and Methods. (B) HFFs were infected with 30 PFU/cell AdEasyE1 (wild type [WT]) or AdEasyE1Sub19 (S19) for 36 h or mock infected (mock), and the E1B 55-kDa protein was visualized by immunofluorescence as described in Materials and Methods. DAPI, 4′,6-diamidino-2-phenylindole. (C) HFFs were treated with IFN or untreated (BSA) and infected with 3 PFU/cell of the virus indicated for 2 or 72 h. Viral DNA concentrations were measured by quantitative PCR, with cellular β-actin DNA used as an internal control, as described in Materials and Methods, and are expressed relative to the input (2 h) concentrations. The values represent the averages of two independent experiments, with cumulative standard deviations indicated by the error bars.

    Journal: Journal of Virology

    Article Title: The Repression Domain of the E1B 55-Kilodalton Protein Participates in Countering Interferon-Induced Inhibition of Adenovirus Replication

    doi: 10.1128/JVI.03387-12

    Figure Lengend Snippet: Viral DNA synthesis is inhibited in AdEasyE1Sub19-infected cells treated with IFN. (A) HFFs were infected with 30 PFU/cell AdEasyE1 (wild type [WT]), AdEasyE1Δ2347 (ΔE1B), or AdEasyE1Sub19 (S19) for the periods indicated or mock infected (M). The proteins listed at the right were examined by immunoblotting of total cell lysates as described in Materials and Methods. (B) HFFs were infected with 30 PFU/cell AdEasyE1 (wild type [WT]) or AdEasyE1Sub19 (S19) for 36 h or mock infected (mock), and the E1B 55-kDa protein was visualized by immunofluorescence as described in Materials and Methods. DAPI, 4′,6-diamidino-2-phenylindole. (C) HFFs were treated with IFN or untreated (BSA) and infected with 3 PFU/cell of the virus indicated for 2 or 72 h. Viral DNA concentrations were measured by quantitative PCR, with cellular β-actin DNA used as an internal control, as described in Materials and Methods, and are expressed relative to the input (2 h) concentrations. The values represent the averages of two independent experiments, with cumulative standard deviations indicated by the error bars.

    Article Snippet: Following initial screening by cleavage of the products of mutagenesis with the appropriate restriction endonuclease, the presence of the desired mutation(s) and the absence of other changes in the E1B gene were confirmed by sequencing (Genewiz).

    Techniques: DNA Synthesis, Infection, Immunofluorescence, Real-time Polymerase Chain Reaction

    Effects of the E1B 55-kDa Sub19 alterations on activation of Stat1. Mock-infected HFFs were treated with 250 units/ml IFN or vehicle-only control (BSA) for 24 h. Cells were infected with 200 PFU/cell AdEasyE1 (wild type [WT]), AdEasyE1Δ2347 (ΔE1B), or AdEasyE1Sub19 (S19) for the periods indicated, and the proteins listed at the right were examined by immunoblotting of whole-cell lysates as described in Materials and Methods. Stat1 phosphorylated on Tyr701 is designated Stat1-P.

    Journal: Journal of Virology

    Article Title: The Repression Domain of the E1B 55-Kilodalton Protein Participates in Countering Interferon-Induced Inhibition of Adenovirus Replication

    doi: 10.1128/JVI.03387-12

    Figure Lengend Snippet: Effects of the E1B 55-kDa Sub19 alterations on activation of Stat1. Mock-infected HFFs were treated with 250 units/ml IFN or vehicle-only control (BSA) for 24 h. Cells were infected with 200 PFU/cell AdEasyE1 (wild type [WT]), AdEasyE1Δ2347 (ΔE1B), or AdEasyE1Sub19 (S19) for the periods indicated, and the proteins listed at the right were examined by immunoblotting of whole-cell lysates as described in Materials and Methods. Stat1 phosphorylated on Tyr701 is designated Stat1-P.

    Article Snippet: Following initial screening by cleavage of the products of mutagenesis with the appropriate restriction endonuclease, the presence of the desired mutation(s) and the absence of other changes in the E1B gene were confirmed by sequencing (Genewiz).

    Techniques: Activation Assay, Infection

    The Sub19 substitutions impair repression of expression of IFN-inducible genes by the E1B 55-kDa protein. HFFs untreated (BSA) or treated with IFN (IFN) were infected with 200 PFU/cell AdEasyE1 (wild type [WT]), AdEasyE1Δ2347 (ΔE1B), or AdEasyE1Sub19 (S19), and total cell RNA was isolated 47 h after infection. The concentrations of GBP1 (A) and IFIT2 (B) mRNAs were determined by quantitative PCR, after synthesis of cDNA by reverse transcription from random primers, as described in Materials and Methods. The values shown represent the averages and cumulative standard deviations (bars) of two independent experiments.

    Journal: Journal of Virology

    Article Title: The Repression Domain of the E1B 55-Kilodalton Protein Participates in Countering Interferon-Induced Inhibition of Adenovirus Replication

    doi: 10.1128/JVI.03387-12

    Figure Lengend Snippet: The Sub19 substitutions impair repression of expression of IFN-inducible genes by the E1B 55-kDa protein. HFFs untreated (BSA) or treated with IFN (IFN) were infected with 200 PFU/cell AdEasyE1 (wild type [WT]), AdEasyE1Δ2347 (ΔE1B), or AdEasyE1Sub19 (S19), and total cell RNA was isolated 47 h after infection. The concentrations of GBP1 (A) and IFIT2 (B) mRNAs were determined by quantitative PCR, after synthesis of cDNA by reverse transcription from random primers, as described in Materials and Methods. The values shown represent the averages and cumulative standard deviations (bars) of two independent experiments.

    Article Snippet: Following initial screening by cleavage of the products of mutagenesis with the appropriate restriction endonuclease, the presence of the desired mutation(s) and the absence of other changes in the E1B gene were confirmed by sequencing (Genewiz).

    Techniques: Expressing, Infection, Isolation, Real-time Polymerase Chain Reaction

    Phenotypic characterization of a stable transgenic line. ( a ) Graphic representation of the DNA constructs used for transgenesis and breeding. ( b ) A 24 h.p.f. embryo expresses Smoa1-EGFP in the forebrain and retinal neuroblasts. A 96 h.p.f. double transgenic embryo derived from crossing of the Tg(krt5:Gal4VP16;14 × UAS:smoa1-EGFP) and the Tg(UAS-E1b:nfsB-mCherry) c264 stable lines expressed GFP ( c ) and mCherry ( d ) in the ONs (arrows). ( e ) A 6-month-old wild-type and ( f ) a transgenic fish with a gross tumor in its left eye (arrow). ( g ) Wild-type fish formed a normal optic chiasm, whereas ( h ) transgenic fish failed to form optic chiasm and showed dramatic expansion of the ONH region (arrow). ( i ) Dissected brain and eyes showing an enlarged ON associating with a gross eye tumor (arrow).

    Journal: Oncogenesis

    Article Title: Activation of Sonic hedgehog signaling in neural progenitor cells promotes glioma development in the zebrafish optic pathway

    doi: 10.1038/oncsis.2014.10

    Figure Lengend Snippet: Phenotypic characterization of a stable transgenic line. ( a ) Graphic representation of the DNA constructs used for transgenesis and breeding. ( b ) A 24 h.p.f. embryo expresses Smoa1-EGFP in the forebrain and retinal neuroblasts. A 96 h.p.f. double transgenic embryo derived from crossing of the Tg(krt5:Gal4VP16;14 × UAS:smoa1-EGFP) and the Tg(UAS-E1b:nfsB-mCherry) c264 stable lines expressed GFP ( c ) and mCherry ( d ) in the ONs (arrows). ( e ) A 6-month-old wild-type and ( f ) a transgenic fish with a gross tumor in its left eye (arrow). ( g ) Wild-type fish formed a normal optic chiasm, whereas ( h ) transgenic fish failed to form optic chiasm and showed dramatic expansion of the ONH region (arrow). ( i ) Dissected brain and eyes showing an enlarged ON associating with a gross eye tumor (arrow).

    Article Snippet: Zebrafish husbandry AB and Tg(UAS-E1b:nfsB-mCherry) c264 strains were acquired from the Zebrafish International Resource Center (ZIRC, Eugene, OR, USA).

    Techniques: Transgenic Assay, Construct, Derivative Assay, Fluorescence In Situ Hybridization

    KSHV ORF57 interacts with PABPC1. (A) ORF57 interacts with PABPC1, but not with E1B-AP5, as shown by coimmunoprecipitation assays. Total cell extracts from TREx BCBL1-Rta cells induced by Dox (1 μg/ml) and treated with a mixture of RNase A and T1 were immunoprecipitated with preimmune mouse IgG- or monoclonal anti-ORF57 antibody-coated beads and examined for PABPC1 and E1B-AP5 by Western blotting (left). Conversely, the same extracts treated with a mixture of RNase A and T1 were immunoprecipitated with nonspecific rabbit IgG- or polyclonal anti-PABPC1 or anti-E1B-AP5 antibody-coated beads and blotted for ORF57 by Western blotting (right). (B) PABPC1 directly interacts with ORF57, as shown by GST pulldown assays. Full-length PABPC1 protein with an N-terminal GST tag or untagged GST was immobilized on glutathione-Sepharose 4B beads and incubated at 4°C overnight in IP buffer complemented with KSHV ORF57 protein containing a C-terminal FLAG tag that was expressed from baculovirus. The glutathione-Sepharose 4B beads without any GST protein were used as an additional negative control. After binding, the beads were washed five times with IP buffer, resuspended in 2× SDS sample buffer, and analyzed by Western blotting with two antibodies, an anti-GST antibody recognizing PABPC1 and an anti-FLAG antibody that recognizes the epitope-tagged ORF57.

    Journal: Journal of Virology

    Article Title: Interplay between Polyadenylate-Binding Protein 1 and Kaposi's Sarcoma-Associated Herpesvirus ORF57 in Accumulation of Polyadenylated Nuclear RNA, a Viral Long Noncoding RNA

    doi: 10.1128/JVI.01693-12

    Figure Lengend Snippet: KSHV ORF57 interacts with PABPC1. (A) ORF57 interacts with PABPC1, but not with E1B-AP5, as shown by coimmunoprecipitation assays. Total cell extracts from TREx BCBL1-Rta cells induced by Dox (1 μg/ml) and treated with a mixture of RNase A and T1 were immunoprecipitated with preimmune mouse IgG- or monoclonal anti-ORF57 antibody-coated beads and examined for PABPC1 and E1B-AP5 by Western blotting (left). Conversely, the same extracts treated with a mixture of RNase A and T1 were immunoprecipitated with nonspecific rabbit IgG- or polyclonal anti-PABPC1 or anti-E1B-AP5 antibody-coated beads and blotted for ORF57 by Western blotting (right). (B) PABPC1 directly interacts with ORF57, as shown by GST pulldown assays. Full-length PABPC1 protein with an N-terminal GST tag or untagged GST was immobilized on glutathione-Sepharose 4B beads and incubated at 4°C overnight in IP buffer complemented with KSHV ORF57 protein containing a C-terminal FLAG tag that was expressed from baculovirus. The glutathione-Sepharose 4B beads without any GST protein were used as an additional negative control. After binding, the beads were washed five times with IP buffer, resuspended in 2× SDS sample buffer, and analyzed by Western blotting with two antibodies, an anti-GST antibody recognizing PABPC1 and an anti-FLAG antibody that recognizes the epitope-tagged ORF57.

    Article Snippet: Rabbit anti-PABPC1 antibody (ab21060; Abcam) and anti-E1B-AP5 antibody (ab68480; Abcam) were used for IP.

    Techniques: Immunoprecipitation, Western Blot, Incubation, FLAG-tag, Negative Control, Binding Assay

    ). This model proposes that the PAN MRE-II-mediated interactions and cross-talks between 5′ capping and 3′ polyadenylation machineries via ORF57, PABPC1, and E1B-AP5 maximize PAN stability.

    Journal: Journal of Virology

    Article Title: Interplay between Polyadenylate-Binding Protein 1 and Kaposi's Sarcoma-Associated Herpesvirus ORF57 in Accumulation of Polyadenylated Nuclear RNA, a Viral Long Noncoding RNA

    doi: 10.1128/JVI.01693-12

    Figure Lengend Snippet: ). This model proposes that the PAN MRE-II-mediated interactions and cross-talks between 5′ capping and 3′ polyadenylation machineries via ORF57, PABPC1, and E1B-AP5 maximize PAN stability.

    Article Snippet: Rabbit anti-PABPC1 antibody (ab21060; Abcam) and anti-E1B-AP5 antibody (ab68480; Abcam) were used for IP.

    Techniques:

    KSHV lytic infection induces host gene shutoff and nuclear distribution of cytoplasmic PABPC1. (A) Pie chart summary of PABPC1 status in ORF57-expressing BCBL1 cells after KSHV lytic induction. TREx BCBL1-Rta cells induced with Dox for 48 h were fixed and double stained with anti-PABPC1 and anti-ORF57 antibodies and imaged by confocal microscopy. A total of 282 cells, with or without PABPC1 and ORF57, were randomly counted in multiple microscope image fields, and only 209 cells with ORF57 staining were calculated for PABPC1 status. (B) Reduction of PABPC1 and β-tubulin in BCBL1 cells with lytic KSHV induction. Total protein was prepared from BCBL1 cells 48 h after lytic KSHV infection induced by 1 mM VA and host gene expression of PABPC1 and β-tubulin, and the presence of ORF57 was examined by Western blotting. BCBL1 cells without lytic induction (uninduced [unind.]) served as a control. (C and D) ORF57-expressing B cells undergoing lytic KSHV infection exhibit redistribution of PABPC1 but not E1B-AP5. TREx BCBL1-vector (a to d) or TREx BCBL1-Rta (e to h) cells induced by Dox were stained with either rabbit anti-PABPC1 (C) or E1B-AP5 (D) antibodies along with a mouse monoclonal anti-ORF57 antibody. Confocal imaging was performed as described in Materials and Methods. The nuclear distributions of cytoplasmic PABPC1 (red) (C) and nuclear E1B-AP5 (red) (D) were seen in ORF57-positive (green) TREx BCBL1-Rta cells. DAPI, 4′,6′-diamino-2-phenylindole nuclear staining.

    Journal: Journal of Virology

    Article Title: Interplay between Polyadenylate-Binding Protein 1 and Kaposi's Sarcoma-Associated Herpesvirus ORF57 in Accumulation of Polyadenylated Nuclear RNA, a Viral Long Noncoding RNA

    doi: 10.1128/JVI.01693-12

    Figure Lengend Snippet: KSHV lytic infection induces host gene shutoff and nuclear distribution of cytoplasmic PABPC1. (A) Pie chart summary of PABPC1 status in ORF57-expressing BCBL1 cells after KSHV lytic induction. TREx BCBL1-Rta cells induced with Dox for 48 h were fixed and double stained with anti-PABPC1 and anti-ORF57 antibodies and imaged by confocal microscopy. A total of 282 cells, with or without PABPC1 and ORF57, were randomly counted in multiple microscope image fields, and only 209 cells with ORF57 staining were calculated for PABPC1 status. (B) Reduction of PABPC1 and β-tubulin in BCBL1 cells with lytic KSHV induction. Total protein was prepared from BCBL1 cells 48 h after lytic KSHV infection induced by 1 mM VA and host gene expression of PABPC1 and β-tubulin, and the presence of ORF57 was examined by Western blotting. BCBL1 cells without lytic induction (uninduced [unind.]) served as a control. (C and D) ORF57-expressing B cells undergoing lytic KSHV infection exhibit redistribution of PABPC1 but not E1B-AP5. TREx BCBL1-vector (a to d) or TREx BCBL1-Rta (e to h) cells induced by Dox were stained with either rabbit anti-PABPC1 (C) or E1B-AP5 (D) antibodies along with a mouse monoclonal anti-ORF57 antibody. Confocal imaging was performed as described in Materials and Methods. The nuclear distributions of cytoplasmic PABPC1 (red) (C) and nuclear E1B-AP5 (red) (D) were seen in ORF57-positive (green) TREx BCBL1-Rta cells. DAPI, 4′,6′-diamino-2-phenylindole nuclear staining.

    Article Snippet: Rabbit anti-PABPC1 antibody (ab21060; Abcam) and anti-E1B-AP5 antibody (ab68480; Abcam) were used for IP.

    Techniques: Infection, Expressing, Staining, Confocal Microscopy, Microscopy, Western Blot, Plasmid Preparation, Imaging

    Roles of PABPC1 and E1B-AP5 in PAN expression. (A) Endogenous PABPC1 is inhibitory for PAN expression. PABPC1 in HeLa cells was knocked down using a PABPC1-specific siRNA (siPABPC1) in the presence or absence of ORF57. A nonspecific siRNA (siControl) was employed as a control. The PABPC1-knockdown efficiency was analyzed by Western blotting (WB). Total RNA of HeLa cells transfected with siPABPC1 or siControl, together with PAN-expressing vector pJM1 in the presence (+) or absence (−) of ORF57, was examined by Northern blotting (NB) with a 32 ) were transfected twice, at intervals of 24 h, with 40 nM PABPC1-specific siRNA or a nonspecific control siRNA before viral lytic induction by either 3 mM butyrate for 24 h (Bu 24) or 1 mM valproate for 48 h (VA 48). The cells were harvested after virus induction for total protein and RNA preparation. The efficiency of PABPC1 knockdown was examined by Western blotting of total protein, and β-tubulin served as a loading control. PAN RNA was determined by Northern blotting of 3 μg of total RNA for Bac36-wt cells and 5 μg of total RNA for Bac36-Δ57 cells using the 32 P-labeled, PAN-specific oligonucleotide probe oJM7. GAPDH RNA in panels A to C served as a sample loading control. The relative levels (signal intensity) of PAN RNA in each sample in panel C were calculated after normalization to the level of GAPDH RNA. A relative ratio (fold) of PAN in siPABPC1- to siControl-treated cells in panels A to C was determined according to the normalized levels in each sample pair.

    Journal: Journal of Virology

    Article Title: Interplay between Polyadenylate-Binding Protein 1 and Kaposi's Sarcoma-Associated Herpesvirus ORF57 in Accumulation of Polyadenylated Nuclear RNA, a Viral Long Noncoding RNA

    doi: 10.1128/JVI.01693-12

    Figure Lengend Snippet: Roles of PABPC1 and E1B-AP5 in PAN expression. (A) Endogenous PABPC1 is inhibitory for PAN expression. PABPC1 in HeLa cells was knocked down using a PABPC1-specific siRNA (siPABPC1) in the presence or absence of ORF57. A nonspecific siRNA (siControl) was employed as a control. The PABPC1-knockdown efficiency was analyzed by Western blotting (WB). Total RNA of HeLa cells transfected with siPABPC1 or siControl, together with PAN-expressing vector pJM1 in the presence (+) or absence (−) of ORF57, was examined by Northern blotting (NB) with a 32 ) were transfected twice, at intervals of 24 h, with 40 nM PABPC1-specific siRNA or a nonspecific control siRNA before viral lytic induction by either 3 mM butyrate for 24 h (Bu 24) or 1 mM valproate for 48 h (VA 48). The cells were harvested after virus induction for total protein and RNA preparation. The efficiency of PABPC1 knockdown was examined by Western blotting of total protein, and β-tubulin served as a loading control. PAN RNA was determined by Northern blotting of 3 μg of total RNA for Bac36-wt cells and 5 μg of total RNA for Bac36-Δ57 cells using the 32 P-labeled, PAN-specific oligonucleotide probe oJM7. GAPDH RNA in panels A to C served as a sample loading control. The relative levels (signal intensity) of PAN RNA in each sample in panel C were calculated after normalization to the level of GAPDH RNA. A relative ratio (fold) of PAN in siPABPC1- to siControl-treated cells in panels A to C was determined according to the normalized levels in each sample pair.

    Article Snippet: Rabbit anti-PABPC1 antibody (ab21060; Abcam) and anti-E1B-AP5 antibody (ab68480; Abcam) were used for IP.

    Techniques: Expressing, Western Blot, Transfection, Plasmid Preparation, Northern Blot, Labeling

    HCMV IE1 and IE2 genes and their products are not present in transformed BRK cell lines. ( A ) Lack of expression of the IE1 or IE2 protein in transformed BRK cell lines when assayed by immunoprecipitation of 35 S-labeled cell extracts using monoclonal antibody mAB810. A-1 is a cell line established by transfection with the E1A-expressing plasmid alone. A/1/2–1 to A/1/2–10 are cell lines transformed by E1A plus IE1 and IE2. Lanes designated HCMV INF display immunoprecipitates from HCMV-infected human foreskin fibroblasts; lane 13 is an exposure 1/8 as long as that presented for lanes 1–12. Numbers to the left of the gel mark the positions of marker proteins whose sizes are indicated in kDa. ( B ) Southern blot analysis showing the lack of IE1 - or IE2 -specific DNA in E1A/IE1/IE2-induced transformants (lanes 1–10). A/B-1 and A/B-2 are cell lines transformed with E1A and E1B-19-kDa. HeLa, 293, and HP1.14 (a derivative HeLa cells expressing IE1) are included as controls. pCGN-IE1 and pCGN-IE2 are IE1 and IE2 expression plasmids, respectively, in amounts equivalent to the rest of the samples, assuming they contain one copy of each of the HCMV genes per cell. Numbers to the left of the gel are marker DNA sizes in kb. ( C ) Expression of E1A protein as determined by immunoprecipitation followed by immunoblot analysis, both with monoclonal antibody M73 to E1A. Lanes are labeled as in A and B . Numbers to the left of the gel are marker protein sizes in kDa. Autoradiograms were scanned and cropped using Photoshop and figures were prepared using Freehand software.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Human cytomegalovirus IE1 and IE2 proteins are mutagenic and mediate "hit-and-run" oncogenic transformation in cooperation with the adenovirus E1A proteins

    doi:

    Figure Lengend Snippet: HCMV IE1 and IE2 genes and their products are not present in transformed BRK cell lines. ( A ) Lack of expression of the IE1 or IE2 protein in transformed BRK cell lines when assayed by immunoprecipitation of 35 S-labeled cell extracts using monoclonal antibody mAB810. A-1 is a cell line established by transfection with the E1A-expressing plasmid alone. A/1/2–1 to A/1/2–10 are cell lines transformed by E1A plus IE1 and IE2. Lanes designated HCMV INF display immunoprecipitates from HCMV-infected human foreskin fibroblasts; lane 13 is an exposure 1/8 as long as that presented for lanes 1–12. Numbers to the left of the gel mark the positions of marker proteins whose sizes are indicated in kDa. ( B ) Southern blot analysis showing the lack of IE1 - or IE2 -specific DNA in E1A/IE1/IE2-induced transformants (lanes 1–10). A/B-1 and A/B-2 are cell lines transformed with E1A and E1B-19-kDa. HeLa, 293, and HP1.14 (a derivative HeLa cells expressing IE1) are included as controls. pCGN-IE1 and pCGN-IE2 are IE1 and IE2 expression plasmids, respectively, in amounts equivalent to the rest of the samples, assuming they contain one copy of each of the HCMV genes per cell. Numbers to the left of the gel are marker DNA sizes in kb. ( C ) Expression of E1A protein as determined by immunoprecipitation followed by immunoblot analysis, both with monoclonal antibody M73 to E1A. Lanes are labeled as in A and B . Numbers to the left of the gel are marker protein sizes in kDa. Autoradiograms were scanned and cropped using Photoshop and figures were prepared using Freehand software.

    Article Snippet: For the two sets of coverslips collected from cells transfected with vectors expressing the E1A plus E1B-19-kDa proteins, one set was stained with anti-E1A antibody (M73), and the other was stained with monoclonal antibody to E1B-19-kDa (Oncogene Science).

    Techniques: Transformation Assay, Expressing, Immunoprecipitation, Labeling, Transfection, Plasmid Preparation, Infection, Marker, Southern Blot, Software

    The IE1 and IE2 gene products are expressed transiently during the transformation of BRK cells. BRK cells seeded on coverslips were transfected with expression plasmids for E1A and E1B-19-kDa ( Left ) or plasmids for E1A and IE1 and IE2 ( Right ). At 2, 5, 13, and 19 days after transfection, two coverslips from each transfection were fixed and kept in phosphate-buffered saline until coverslips for all time points were collected. Expression of the transfected genes was examined by indirect immunofluorescent staining with appropriate monoclonal antibodies. Anti-E1A, anti-E1B-19-kDa, and anti-IE1/2 identify the antibodies used for the immunofluorescent staining. Images are from independent coverslips. The images from days 13 and 19 display transformed foci whose cells were positive for E1A and E1B-19-kDa proteins, but completely negative for IE1 and IE2 protein expression. (×50).

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Human cytomegalovirus IE1 and IE2 proteins are mutagenic and mediate "hit-and-run" oncogenic transformation in cooperation with the adenovirus E1A proteins

    doi:

    Figure Lengend Snippet: The IE1 and IE2 gene products are expressed transiently during the transformation of BRK cells. BRK cells seeded on coverslips were transfected with expression plasmids for E1A and E1B-19-kDa ( Left ) or plasmids for E1A and IE1 and IE2 ( Right ). At 2, 5, 13, and 19 days after transfection, two coverslips from each transfection were fixed and kept in phosphate-buffered saline until coverslips for all time points were collected. Expression of the transfected genes was examined by indirect immunofluorescent staining with appropriate monoclonal antibodies. Anti-E1A, anti-E1B-19-kDa, and anti-IE1/2 identify the antibodies used for the immunofluorescent staining. Images are from independent coverslips. The images from days 13 and 19 display transformed foci whose cells were positive for E1A and E1B-19-kDa proteins, but completely negative for IE1 and IE2 protein expression. (×50).

    Article Snippet: For the two sets of coverslips collected from cells transfected with vectors expressing the E1A plus E1B-19-kDa proteins, one set was stained with anti-E1A antibody (M73), and the other was stained with monoclonal antibody to E1B-19-kDa (Oncogene Science).

    Techniques: Transformation Assay, Transfection, Expressing, Staining

    Viral DNA synthesis is inhibited in AdEasyE1Sub19-infected cells treated with IFN. (A) HFFs were infected with 30 PFU/cell AdEasyE1 (wild type [WT]), AdEasyE1Δ2347 (ΔE1B), or AdEasyE1Sub19 (S19) for the periods indicated or mock infected (M). The proteins listed at the right were examined by immunoblotting of total cell lysates as described in Materials and Methods. (B) HFFs were infected with 30 PFU/cell AdEasyE1 (wild type [WT]) or AdEasyE1Sub19 (S19) for 36 h or mock infected (mock), and the E1B 55-kDa protein was visualized by immunofluorescence as described in Materials and Methods. DAPI, 4′,6-diamidino-2-phenylindole. (C) HFFs were treated with IFN or untreated (BSA) and infected with 3 PFU/cell of the virus indicated for 2 or 72 h. Viral DNA concentrations were measured by quantitative PCR, with cellular β-actin DNA used as an internal control, as described in Materials and Methods, and are expressed relative to the input (2 h) concentrations. The values represent the averages of two independent experiments, with cumulative standard deviations indicated by the error bars.

    Journal: Journal of Virology

    Article Title: The Repression Domain of the E1B 55-Kilodalton Protein Participates in Countering Interferon-Induced Inhibition of Adenovirus Replication

    doi: 10.1128/JVI.03387-12

    Figure Lengend Snippet: Viral DNA synthesis is inhibited in AdEasyE1Sub19-infected cells treated with IFN. (A) HFFs were infected with 30 PFU/cell AdEasyE1 (wild type [WT]), AdEasyE1Δ2347 (ΔE1B), or AdEasyE1Sub19 (S19) for the periods indicated or mock infected (M). The proteins listed at the right were examined by immunoblotting of total cell lysates as described in Materials and Methods. (B) HFFs were infected with 30 PFU/cell AdEasyE1 (wild type [WT]) or AdEasyE1Sub19 (S19) for 36 h or mock infected (mock), and the E1B 55-kDa protein was visualized by immunofluorescence as described in Materials and Methods. DAPI, 4′,6-diamidino-2-phenylindole. (C) HFFs were treated with IFN or untreated (BSA) and infected with 3 PFU/cell of the virus indicated for 2 or 72 h. Viral DNA concentrations were measured by quantitative PCR, with cellular β-actin DNA used as an internal control, as described in Materials and Methods, and are expressed relative to the input (2 h) concentrations. The values represent the averages of two independent experiments, with cumulative standard deviations indicated by the error bars.

    Article Snippet: The sequence of the E1B 55-kDa insert was verified by sequencing (Genewiz).

    Techniques: DNA Synthesis, Infection, Immunofluorescence, Real-time Polymerase Chain Reaction

    Effects of the E1B 55-kDa Sub19 alterations on activation of Stat1. Mock-infected HFFs were treated with 250 units/ml IFN or vehicle-only control (BSA) for 24 h. Cells were infected with 200 PFU/cell AdEasyE1 (wild type [WT]), AdEasyE1Δ2347 (ΔE1B), or AdEasyE1Sub19 (S19) for the periods indicated, and the proteins listed at the right were examined by immunoblotting of whole-cell lysates as described in Materials and Methods. Stat1 phosphorylated on Tyr701 is designated Stat1-P.

    Journal: Journal of Virology

    Article Title: The Repression Domain of the E1B 55-Kilodalton Protein Participates in Countering Interferon-Induced Inhibition of Adenovirus Replication

    doi: 10.1128/JVI.03387-12

    Figure Lengend Snippet: Effects of the E1B 55-kDa Sub19 alterations on activation of Stat1. Mock-infected HFFs were treated with 250 units/ml IFN or vehicle-only control (BSA) for 24 h. Cells were infected with 200 PFU/cell AdEasyE1 (wild type [WT]), AdEasyE1Δ2347 (ΔE1B), or AdEasyE1Sub19 (S19) for the periods indicated, and the proteins listed at the right were examined by immunoblotting of whole-cell lysates as described in Materials and Methods. Stat1 phosphorylated on Tyr701 is designated Stat1-P.

    Article Snippet: The sequence of the E1B 55-kDa insert was verified by sequencing (Genewiz).

    Techniques: Activation Assay, Infection

    The Sub19 substitutions impair repression of expression of IFN-inducible genes by the E1B 55-kDa protein. HFFs untreated (BSA) or treated with IFN (IFN) were infected with 200 PFU/cell AdEasyE1 (wild type [WT]), AdEasyE1Δ2347 (ΔE1B), or AdEasyE1Sub19 (S19), and total cell RNA was isolated 47 h after infection. The concentrations of GBP1 (A) and IFIT2 (B) mRNAs were determined by quantitative PCR, after synthesis of cDNA by reverse transcription from random primers, as described in Materials and Methods. The values shown represent the averages and cumulative standard deviations (bars) of two independent experiments.

    Journal: Journal of Virology

    Article Title: The Repression Domain of the E1B 55-Kilodalton Protein Participates in Countering Interferon-Induced Inhibition of Adenovirus Replication

    doi: 10.1128/JVI.03387-12

    Figure Lengend Snippet: The Sub19 substitutions impair repression of expression of IFN-inducible genes by the E1B 55-kDa protein. HFFs untreated (BSA) or treated with IFN (IFN) were infected with 200 PFU/cell AdEasyE1 (wild type [WT]), AdEasyE1Δ2347 (ΔE1B), or AdEasyE1Sub19 (S19), and total cell RNA was isolated 47 h after infection. The concentrations of GBP1 (A) and IFIT2 (B) mRNAs were determined by quantitative PCR, after synthesis of cDNA by reverse transcription from random primers, as described in Materials and Methods. The values shown represent the averages and cumulative standard deviations (bars) of two independent experiments.

    Article Snippet: The sequence of the E1B 55-kDa insert was verified by sequencing (Genewiz).

    Techniques: Expressing, Infection, Isolation, Real-time Polymerase Chain Reaction

    E4-ORF3 induces SUMO conjugation to E1B-55K, Nbs1, and Mre11. (A) HeLa cells infected with dl 355 (ΔE4-ORF6) or dl 355- in ORF3 (ΔE4-ORF3/ΔE4-ORF6) were immunostained for E1B-55K and E2-DBP or E4-ORF3 at 12 hpi. (B) His 6 -tagged SUMO-1 or SUMO-3-expressing HeLa cells were infected with indicating amounts of dl 355 or dl 355- in ORF3. At 12 hpi, SUMO conjugates were captured by Ni-NTA agarose beads and analyzed by Western blotting with the anti-E1B-55K antibody. SUMO-conjugated RanGAP1 was shown as a control for SUMO capture. Protein levels of E1B-55K and E4-ORF3 from cell lysates were determined by Western blotting. (C) GST-tagged Ad5 E1B-55K was incubated with SUMO-1/E1/E2 and the indicated concentrations of Ad5 E4-ORF3. The levels of SUMO conjugation were analyzed by Western blotting with the anti-E1B-55K antibody. (D) GST-tagged Nbs1 and Mre11 were incubated with SUMO E1/E2 and SUMO-1 or SUMO-3 in the absence or presence of Ad5 E4-ORF3 (3 μM). The reaction mixture was analyzed by Western blotting with anti-Nbs1 and anti-Mre11 antibodies.

    Journal: mBio

    Article Title: Mechanism of Adenovirus E4-ORF3-Mediated SUMO Modifications

    doi: 10.1128/mBio.00022-19

    Figure Lengend Snippet: E4-ORF3 induces SUMO conjugation to E1B-55K, Nbs1, and Mre11. (A) HeLa cells infected with dl 355 (ΔE4-ORF6) or dl 355- in ORF3 (ΔE4-ORF3/ΔE4-ORF6) were immunostained for E1B-55K and E2-DBP or E4-ORF3 at 12 hpi. (B) His 6 -tagged SUMO-1 or SUMO-3-expressing HeLa cells were infected with indicating amounts of dl 355 or dl 355- in ORF3. At 12 hpi, SUMO conjugates were captured by Ni-NTA agarose beads and analyzed by Western blotting with the anti-E1B-55K antibody. SUMO-conjugated RanGAP1 was shown as a control for SUMO capture. Protein levels of E1B-55K and E4-ORF3 from cell lysates were determined by Western blotting. (C) GST-tagged Ad5 E1B-55K was incubated with SUMO-1/E1/E2 and the indicated concentrations of Ad5 E4-ORF3. The levels of SUMO conjugation were analyzed by Western blotting with the anti-E1B-55K antibody. (D) GST-tagged Nbs1 and Mre11 were incubated with SUMO E1/E2 and SUMO-1 or SUMO-3 in the absence or presence of Ad5 E4-ORF3 (3 μM). The reaction mixture was analyzed by Western blotting with anti-Nbs1 and anti-Mre11 antibodies.

    Article Snippet: The optimized E1B-55K coding region was synthesized from Biomatik and cloned in pGEX-KG.

    Techniques: Conjugation Assay, Infection, Expressing, Western Blot, Incubation

    Fusion of human cells to mouse A9 cells expressing the E1B-55K and E4orf6 genes permits the E1B-55K protein to enter mouse A9 cell nuclei. Expression of the E1B-55K and E4orf6 proteins was established in mouse A9 cells with the recombinant vaccinia virus T7 infection-transfection system. HeLa cells were harvested by trypsin treatment and allowed to attach to the substrate with adherent A9 cells. The HeLa and A9 cells were fused 8 h after transfection with PEG. The resulting heterokaryons were maintained in the presence of cycloheximide (100 μg/ml) for 4 h before processing for double-label immunofluorescence. (A) Phase contrast image showing two heterokaryons; (B) DAPI-stained image showing DNA; (C) staining for the large subunit of the human Ku protein specifically labels the HeLa cell nuclei; (D) staining for the E1B-55K protein with the rat monoclonal antibody 9C10. A9 cell nuclei are marked by arrowheads.

    Journal: Journal of Virology

    Article Title: An Activity Associated with Human Chromosome 21 Permits Nuclear Colocalization of the Adenovirus E1B-55K and E4orf6 Proteins and Promotes Viral Late Gene Expression

    doi: 10.1128/JVI.77.14.8087-8098.2003

    Figure Lengend Snippet: Fusion of human cells to mouse A9 cells expressing the E1B-55K and E4orf6 genes permits the E1B-55K protein to enter mouse A9 cell nuclei. Expression of the E1B-55K and E4orf6 proteins was established in mouse A9 cells with the recombinant vaccinia virus T7 infection-transfection system. HeLa cells were harvested by trypsin treatment and allowed to attach to the substrate with adherent A9 cells. The HeLa and A9 cells were fused 8 h after transfection with PEG. The resulting heterokaryons were maintained in the presence of cycloheximide (100 μg/ml) for 4 h before processing for double-label immunofluorescence. (A) Phase contrast image showing two heterokaryons; (B) DAPI-stained image showing DNA; (C) staining for the large subunit of the human Ku protein specifically labels the HeLa cell nuclei; (D) staining for the E1B-55K protein with the rat monoclonal antibody 9C10. A9 cell nuclei are marked by arrowheads.

    Article Snippet: Double-label immunofluorescence was performed with the E4orf6 protein-specific mouse monoclonal antibody MAb3 ( ) as undiluted hybridoma culture supernatant fluid and the E1B-55K protein-specific rat monoclonal antibody 9C10 ( ) at 1 μg per ml (Oncogene Science, Uniondale, N.Y.).

    Techniques: Expressing, Recombinant, Infection, Transfection, Immunofluorescence, Staining

    The E1B-55K and E4orf6 proteins colocalize in the nuclei of HeLa cells but not in mouse cells. HeLa cells (A through H) or mouse A9 cells (I through P) were infected with the recombinant vaccinia virus vTF7.3 to establish expression of the T7 RNA polymerase and were then transfected with cDNA under the control of the T7 promoter to express the E1B-55K gene (A through D, I through L) or to express both the E1B-55K and E4orf6 genes (E through H, M through P). Simultaneous double-label immunofluorescence was performed at 15 h after transfection, and representative cells are shown. (A, E, I, and M) DIC image; (B, F, J, and N) staining for the E4orf6 protein with mouse monoclonal antibody MAb3; (C, G, K, and O) staining for the E1B-55K protein with rat monoclonal antibody 9C10; (D, H, L, and P) merged red and green image. The localization of the proteins was recorded by using confocal laser scanning microscopy. Bars, 10 μm.

    Journal: Journal of Virology

    Article Title: An Activity Associated with Human Chromosome 21 Permits Nuclear Colocalization of the Adenovirus E1B-55K and E4orf6 Proteins and Promotes Viral Late Gene Expression

    doi: 10.1128/JVI.77.14.8087-8098.2003

    Figure Lengend Snippet: The E1B-55K and E4orf6 proteins colocalize in the nuclei of HeLa cells but not in mouse cells. HeLa cells (A through H) or mouse A9 cells (I through P) were infected with the recombinant vaccinia virus vTF7.3 to establish expression of the T7 RNA polymerase and were then transfected with cDNA under the control of the T7 promoter to express the E1B-55K gene (A through D, I through L) or to express both the E1B-55K and E4orf6 genes (E through H, M through P). Simultaneous double-label immunofluorescence was performed at 15 h after transfection, and representative cells are shown. (A, E, I, and M) DIC image; (B, F, J, and N) staining for the E4orf6 protein with mouse monoclonal antibody MAb3; (C, G, K, and O) staining for the E1B-55K protein with rat monoclonal antibody 9C10; (D, H, L, and P) merged red and green image. The localization of the proteins was recorded by using confocal laser scanning microscopy. Bars, 10 μm.

    Article Snippet: Double-label immunofluorescence was performed with the E4orf6 protein-specific mouse monoclonal antibody MAb3 ( ) as undiluted hybridoma culture supernatant fluid and the E1B-55K protein-specific rat monoclonal antibody 9C10 ( ) at 1 μg per ml (Oncogene Science, Uniondale, N.Y.).

    Techniques: Infection, Recombinant, Expressing, Transfection, Immunofluorescence, Staining, Confocal Laser Scanning Microscopy

    (A) Localization of E1B-AP5, ATRIP, and RPA32 in mock-infected interphase A549 cells. Cells were grown on glass coverslips and then treated with preextraction buffer and fixed with 4% (wt/vol) paraformaldehyde as described in Materials and Methods. Antigens were detected using the appropriate antibodies. (B) Localization of pTP, DBP, and RPA32 at viral replication centers in Ad5-infected A549 cells. Twenty-four hours postinfection with 10 PFU/cell wt Ad5, cells were treated with preextraction buffer and then fixed with 4% (wt/vol) paraformaldehyde, whereupon antigens were detected using the appropriate reagents. Colocalization images were recorded using a Zeiss LSM510-Meta laser scanning confocal microscope. Nuclei are stained with DAPI and are shown in blue.

    Journal: Journal of Virology

    Article Title: A Role for E1B-AP5 in ATR Signaling Pathways during Adenovirus Infection

    doi: 10.1128/JVI.00170-08

    Figure Lengend Snippet: (A) Localization of E1B-AP5, ATRIP, and RPA32 in mock-infected interphase A549 cells. Cells were grown on glass coverslips and then treated with preextraction buffer and fixed with 4% (wt/vol) paraformaldehyde as described in Materials and Methods. Antigens were detected using the appropriate antibodies. (B) Localization of pTP, DBP, and RPA32 at viral replication centers in Ad5-infected A549 cells. Twenty-four hours postinfection with 10 PFU/cell wt Ad5, cells were treated with preextraction buffer and then fixed with 4% (wt/vol) paraformaldehyde, whereupon antigens were detected using the appropriate reagents. Colocalization images were recorded using a Zeiss LSM510-Meta laser scanning confocal microscope. Nuclei are stained with DAPI and are shown in blue.

    Article Snippet: Small interfering RNA (siRNA) oligonucleotides targeting ATR and E1B-AP5 were purchased from Ambion, and their sequences were as follows: 5′-GGAAUAUAAUACAGUUGUATT-3′ (ATR) and 5′-GCAGUGGAACCAGUACUAUTT-3′ (E1B-AP5).

    Techniques: Infection, Microscopy, Staining

    Effects of Ad infection on protein levels of E1B-55K binding proteins. A549 cells were infected with 10 PFU per cell of either wt Ad5 (A) or wt Ad12 (B). Cells were harvested at the appropriate times postinfection, and 50 μg protein samples was separated by SDS-PAGE and transferred onto nitrocellulose. Membranes were then Western blotted for E1B-AP5 (i), MRE11 (ii), p53 (iii), E1B-55K (iv), and β-actin (v) using the appropriate antibodies and visualized by enhanced chemiluminescence. (C) Immunofluorescent Zeiss LSM510-Meta confocal microscopic detection of E1B-AP5 protein expression in wt Ad5- and wt Ad12-infected A549 cells. Cells were infected with either 10 PFU/cell wt Ad5 or wt Ad12, subsequently fixed with 4% (wt/vol) paraformaldehyde in PBS, and permeabilized with acetone 24 h postinfection. E1B-AP5 (i and iv), Ad5 DBP (ii), and Ad12 E1B-55K (v) were then visualized using the appropriate antibodies. Nuclei are stained with DAPI and are shown in blue.

    Journal: Journal of Virology

    Article Title: A Role for E1B-AP5 in ATR Signaling Pathways during Adenovirus Infection

    doi: 10.1128/JVI.00170-08

    Figure Lengend Snippet: Effects of Ad infection on protein levels of E1B-55K binding proteins. A549 cells were infected with 10 PFU per cell of either wt Ad5 (A) or wt Ad12 (B). Cells were harvested at the appropriate times postinfection, and 50 μg protein samples was separated by SDS-PAGE and transferred onto nitrocellulose. Membranes were then Western blotted for E1B-AP5 (i), MRE11 (ii), p53 (iii), E1B-55K (iv), and β-actin (v) using the appropriate antibodies and visualized by enhanced chemiluminescence. (C) Immunofluorescent Zeiss LSM510-Meta confocal microscopic detection of E1B-AP5 protein expression in wt Ad5- and wt Ad12-infected A549 cells. Cells were infected with either 10 PFU/cell wt Ad5 or wt Ad12, subsequently fixed with 4% (wt/vol) paraformaldehyde in PBS, and permeabilized with acetone 24 h postinfection. E1B-AP5 (i and iv), Ad5 DBP (ii), and Ad12 E1B-55K (v) were then visualized using the appropriate antibodies. Nuclei are stained with DAPI and are shown in blue.

    Article Snippet: Small interfering RNA (siRNA) oligonucleotides targeting ATR and E1B-AP5 were purchased from Ambion, and their sequences were as follows: 5′-GGAAUAUAAUACAGUUGUATT-3′ (ATR) and 5′-GCAGUGGAACCAGUACUAUTT-3′ (E1B-AP5).

    Techniques: Infection, Binding Assay, SDS Page, Western Blot, Expressing, Staining

    Colocalization of E1B-AP5 and RPA32, and ATRIP and RPA32, at Ad5 (A) and Ad12 (B) replication centers. Cells were infected with 10 PFU/cell of the appropriate virus. Ad5-infected and Ad12-infected cells were treated with preextraction buffer and then fixed with 4% (wt/vol) paraformaldehyde 24 h and 48 h postinfection, respectively. E1B-AP5, RPA32, and ATRIP localizations were visualized using the appropriate antibodies. Colocalization images were recorded using a Zeiss LSM510-Meta confocal microscope. Regions of substantial colocalization are shown as yellow in the merged image. Nuclei are stained with DAPI and are shown in blue.

    Journal: Journal of Virology

    Article Title: A Role for E1B-AP5 in ATR Signaling Pathways during Adenovirus Infection

    doi: 10.1128/JVI.00170-08

    Figure Lengend Snippet: Colocalization of E1B-AP5 and RPA32, and ATRIP and RPA32, at Ad5 (A) and Ad12 (B) replication centers. Cells were infected with 10 PFU/cell of the appropriate virus. Ad5-infected and Ad12-infected cells were treated with preextraction buffer and then fixed with 4% (wt/vol) paraformaldehyde 24 h and 48 h postinfection, respectively. E1B-AP5, RPA32, and ATRIP localizations were visualized using the appropriate antibodies. Colocalization images were recorded using a Zeiss LSM510-Meta confocal microscope. Regions of substantial colocalization are shown as yellow in the merged image. Nuclei are stained with DAPI and are shown in blue.

    Article Snippet: Small interfering RNA (siRNA) oligonucleotides targeting ATR and E1B-AP5 were purchased from Ambion, and their sequences were as follows: 5′-GGAAUAUAAUACAGUUGUATT-3′ (ATR) and 5′-GCAGUGGAACCAGUACUAUTT-3′ (E1B-AP5).

    Techniques: Infection, Microscopy, Staining

    (A) Colocalization of E1B-AP5 with E1B-55K at Ad5 and Ad12 replication centers. Cells were infected with 10 PFU/cell of the appropriate virus. Infected cells were treated with preextraction buffer and then fixed with 4% (wt/vol) paraformaldehyde 24 h postinfection. E1B-AP5 and Ad E1B-55K were visualized using the appropriate antibodies. (B) Recruitment of E1B-AP5, RPA32, and ATRIP to Ad5 replication centers is independent of E1B-55K. Cells were infected with 10 PFU/cell of Ad5 dl 1520 and treated with preextraction buffer and then fixed with 4% (wt/vol) paraformaldehyde 24 h postinfection. E1B-AP5, Ad5 DBP, RPA32, and ATRIP were visualized using the appropriate antibodies. Colocalization images were recorded using a Zeiss LSM510-Meta confocal microscope. Regions of substantial colocalization are shown as yellow in the merged image. Nuclei are stained with DAPI and are shown in blue.

    Journal: Journal of Virology

    Article Title: A Role for E1B-AP5 in ATR Signaling Pathways during Adenovirus Infection

    doi: 10.1128/JVI.00170-08

    Figure Lengend Snippet: (A) Colocalization of E1B-AP5 with E1B-55K at Ad5 and Ad12 replication centers. Cells were infected with 10 PFU/cell of the appropriate virus. Infected cells were treated with preextraction buffer and then fixed with 4% (wt/vol) paraformaldehyde 24 h postinfection. E1B-AP5 and Ad E1B-55K were visualized using the appropriate antibodies. (B) Recruitment of E1B-AP5, RPA32, and ATRIP to Ad5 replication centers is independent of E1B-55K. Cells were infected with 10 PFU/cell of Ad5 dl 1520 and treated with preextraction buffer and then fixed with 4% (wt/vol) paraformaldehyde 24 h postinfection. E1B-AP5, Ad5 DBP, RPA32, and ATRIP were visualized using the appropriate antibodies. Colocalization images were recorded using a Zeiss LSM510-Meta confocal microscope. Regions of substantial colocalization are shown as yellow in the merged image. Nuclei are stained with DAPI and are shown in blue.

    Article Snippet: Small interfering RNA (siRNA) oligonucleotides targeting ATR and E1B-AP5 were purchased from Ambion, and their sequences were as follows: 5′-GGAAUAUAAUACAGUUGUATT-3′ (ATR) and 5′-GCAGUGGAACCAGUACUAUTT-3′ (E1B-AP5).

    Techniques: Infection, Microscopy, Staining

    (A) Ad5 and Ad12 differentially regulate the phosphorylation of RPA32, Rad9, and Smc1 during infection. A549 cells were infected with 10 PFU/cell of either wt Ad5 or wt Ad12. Cells were harvested at the appropriate times postinfection, and 50 μg protein samples was separated by SDS-PAGE. After electrophoretic transfer onto nitrocellulose, membranes were Western blotted for RPA32 (i), RPA32 S4/8 (ii), Rad9 (iii), Smc1-S966 (iv), Smc1 (v), Chk1-S345 (vi), Chk1 (vii), γ-H2AX (viii), and H2AX (ix) using the appropriate antibodies. (B) E1B-AP5 is required for Ad12-induced phosphorylation of RPA32. A549 cells were initially treated with either nonsilencing (non-sil.) siRNA or siRNA oligonucleotides specific for the E1B-AP5 gene. Cells were subsequently infected with either Ad5 or Ad12 (at 10 PFU/cell), and whole-cell lysates were prepared at the appropriate times postinfection. After SDS-PAGE and transfer onto nitrocellulose, membranes were probed for E1B-AP5 (i), RPA32 (ii), RPA32 S4/8 (iii), Smc1-S966 (iv), Smc1 (v), γ-H2AX (vi), and H2AX (vii) with the appropriate antibodies. Antigens were visualized by ECL.

    Journal: Journal of Virology

    Article Title: A Role for E1B-AP5 in ATR Signaling Pathways during Adenovirus Infection

    doi: 10.1128/JVI.00170-08

    Figure Lengend Snippet: (A) Ad5 and Ad12 differentially regulate the phosphorylation of RPA32, Rad9, and Smc1 during infection. A549 cells were infected with 10 PFU/cell of either wt Ad5 or wt Ad12. Cells were harvested at the appropriate times postinfection, and 50 μg protein samples was separated by SDS-PAGE. After electrophoretic transfer onto nitrocellulose, membranes were Western blotted for RPA32 (i), RPA32 S4/8 (ii), Rad9 (iii), Smc1-S966 (iv), Smc1 (v), Chk1-S345 (vi), Chk1 (vii), γ-H2AX (viii), and H2AX (ix) using the appropriate antibodies. (B) E1B-AP5 is required for Ad12-induced phosphorylation of RPA32. A549 cells were initially treated with either nonsilencing (non-sil.) siRNA or siRNA oligonucleotides specific for the E1B-AP5 gene. Cells were subsequently infected with either Ad5 or Ad12 (at 10 PFU/cell), and whole-cell lysates were prepared at the appropriate times postinfection. After SDS-PAGE and transfer onto nitrocellulose, membranes were probed for E1B-AP5 (i), RPA32 (ii), RPA32 S4/8 (iii), Smc1-S966 (iv), Smc1 (v), γ-H2AX (vi), and H2AX (vii) with the appropriate antibodies. Antigens were visualized by ECL.

    Article Snippet: Small interfering RNA (siRNA) oligonucleotides targeting ATR and E1B-AP5 were purchased from Ambion, and their sequences were as follows: 5′-GGAAUAUAAUACAGUUGUATT-3′ (ATR) and 5′-GCAGUGGAACCAGUACUAUTT-3′ (E1B-AP5).

    Techniques: Infection, SDS Page, Western Blot

    Effect of overexpression and siRNA knockdown of NRF2 on E1b expression

    Journal: Biochimica et biophysica acta

    Article Title: Intronic DNA elements regulate Nrf-2 chemical responsiveness of the human microsomal epoxide hydrolase gene (EPHX1) through a far upstream alternative promoter

    doi: 10.1016/j.bbagrm.2014.03.014

    Figure Lengend Snippet: Effect of overexpression and siRNA knockdown of NRF2 on E1b expression

    Article Snippet: Therefore, although the factors contributing to the E1b chromatin looping remain to be elucidated, they represent unique targets as E1b expression modulators that may in turn be regulated through the action of xenobiotics at this distal enhancer.

    Techniques: Over Expression, Expressing

    Identification of regulatory elements involved in E1b induction

    Journal: Biochimica et biophysica acta

    Article Title: Intronic DNA elements regulate Nrf-2 chemical responsiveness of the human microsomal epoxide hydrolase gene (EPHX1) through a far upstream alternative promoter

    doi: 10.1016/j.bbagrm.2014.03.014

    Figure Lengend Snippet: Identification of regulatory elements involved in E1b induction

    Article Snippet: Therefore, although the factors contributing to the E1b chromatin looping remain to be elucidated, they represent unique targets as E1b expression modulators that may in turn be regulated through the action of xenobiotics at this distal enhancer.

    Techniques:

    Role of DNase HS sites in Nrf2-mediated induction of E1b

    Journal: Biochimica et biophysica acta

    Article Title: Intronic DNA elements regulate Nrf-2 chemical responsiveness of the human microsomal epoxide hydrolase gene (EPHX1) through a far upstream alternative promoter

    doi: 10.1016/j.bbagrm.2014.03.014

    Figure Lengend Snippet: Role of DNase HS sites in Nrf2-mediated induction of E1b

    Article Snippet: Therefore, although the factors contributing to the E1b chromatin looping remain to be elucidated, they represent unique targets as E1b expression modulators that may in turn be regulated through the action of xenobiotics at this distal enhancer.

    Techniques:

    Antioxidants induce E1b expression

    Journal: Biochimica et biophysica acta

    Article Title: Intronic DNA elements regulate Nrf-2 chemical responsiveness of the human microsomal epoxide hydrolase gene (EPHX1) through a far upstream alternative promoter

    doi: 10.1016/j.bbagrm.2014.03.014

    Figure Lengend Snippet: Antioxidants induce E1b expression

    Article Snippet: Therefore, although the factors contributing to the E1b chromatin looping remain to be elucidated, they represent unique targets as E1b expression modulators that may in turn be regulated through the action of xenobiotics at this distal enhancer.

    Techniques: Expressing