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  • 99
    Thermo Fisher reverse transcribed rna
    Reverse Transcribed Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 978 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher reverse transcribed
    Reverse Transcribed, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 4103 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    75
    TaKaRa reverse transcribed reagent
    Reverse Transcribed Reagent, supplied by TaKaRa, used in various techniques. Bioz Stars score: 75/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa reverse transcribed kit
    Reverse Transcribed Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 80/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Thermo Fisher reverse transcribed polymerase chain reaction
    Reverse Transcribed Polymerase Chain Reaction, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Roche reverse transcribed cdna
    Reverse Transcribed Cdna, supplied by Roche, used in various techniques. Bioz Stars score: 95/100, based on 145 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Thermo Fisher reverse transcribed superscript invitrogen
    Reverse Transcribed Superscript Invitrogen, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 78/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Boehringer Mannheim reverse transcribed
    Reverse Transcribed, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 88/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Becton Dickinson reverse transcribed
    Reverse Transcribed, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 88/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    TaKaRa reverse transcribed
    Reverse Transcribed, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 723 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    PrimerDesign Inc reverse transcribed cdna
    Reverse Transcribed Cdna, supplied by PrimerDesign Inc, used in various techniques. Bioz Stars score: 91/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Meridian Life Science reverse transcribed
    Reverse Transcribed, supplied by Meridian Life Science, 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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    Thermo Fisher cdna reverse transcribed
    Cdna Reverse Transcribed, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 131 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    GE Healthcare reverse transcribed cdnas
    Reverse Transcribed Cdnas, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 79/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher cdnas reverse transcribed
    Cdnas Reverse Transcribed, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare reverse transcribed probes
    Reverse Transcribed Probes, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 78/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Bio-Rad reverse transcribed iscript rt supermix
    Reverse Transcribed Iscript Rt Supermix, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Diagenode reverse transcribed cdnas
    Reverse Transcribed Cdnas, supplied by Diagenode, used in various techniques. Bioz Stars score: 84/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    Toyobo reverse transcribed kit
    Reverse Transcribed Kit, supplied by Toyobo, used in various techniques. Bioz Stars score: 77/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Bio-Rad spectrophotometry reverse transcribed
    Spectrophotometry Reverse Transcribed, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 84/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa reverse transcribed cdnas
    Reverse Transcribed Cdnas, supplied by TaKaRa, used in various techniques. Bioz Stars score: 89/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher reverse transcribed synthesis amino allyl kit
    Reverse Transcribed Synthesis Amino Allyl Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 84/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Thermo Fisher reverse transcribed pcr buffers
    Reverse Transcribed Pcr Buffers, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 79/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad cdna reverse transcribed
    Cdna Reverse Transcribed, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 82/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Bio-Rad reverse transcribed rna
    Design and expression of peGn, pscDEC-eGn and pscCD11c-eGn. a Schematic representation of peGn, pscDEC-eGn and pscCD11c-eGn. peGn includes the codon-optimized eGn sequence with its intrinsic signal peptide (SP, RVFV M segment from nt 411 to nt 1763) and cloned in the pcDNA4-V5His expression vector. pscDEC-eGn and pscCD11c include the signal peptide (SP) of the VH chain of the parental mAb sequence, the scFv (VH and VL) sequence and the nt 477 to nt 1763 portion of the codon-optimized eGn sequence. The VH and VL sequences are connected together with a (G 4 S) 4 linker and the scFv and eGn sequences are connected with a (G 4 S) 3 linker. These chimeric sequences were cloned in a pcDNA3.1 vector. b, c HEK293 cells were transfected with pcDNA3.1, peGn, pscDEC-eGn and pscCD11c-eGn. In ( b ), the cell lysates (10 µl) were separated by SDS-PAGE under reducing conditions and eGn was revealed with anti-RVFV HMAF followed by a HRP-GAM IgG. The predicted sizes of the expressed untargeted and chimeric eGn proteins are 48.8 kDa for untargeted eGn, 76.3 kDa for the scDEC-eGn protein and 78.6 kDa for the scCD11c-eGn protein. All samples were gathered from the same experiment and were processed in parallel (1 repeated experiment). In ( c ) the cell lysates (100 µl) and the concentrated cell supernatants (500 µl) gathered from the same experiment were spotted onto the same nitrocellulose membrane and the eGn protein was revealed as in ( b ) (1 repeated experiment). d Sheep skin lymph low-density cells were reacted with the parental anti-DEC205, anti-CD11c or isotype control mAbs (top panel) or with the concentrated supernatant from HEK293 cells transfected with peGn, pscDEC-eGn, pscCD11c-eGn and a negative control pcDNA3.1 (bottom panel). The staining of the DAPI neg FSC hi cells, which include dominantly cDCs, is depicted (dark gray) compared to control (dash line). Bound mAbs were revealed with A488-DAM IgG and bound eGn with anti-RVFV HMAF followed by A488-DAM IgG. e Relative expression of peGn, pscDEC-eGn and pscCD11c-eGn in sheep skin. peGn, pscDEC-eGn and pscCD11c-eGn (100 µg) were injected intradermally in 100 µl saline in inner front leg sites of two sheep followed by SEP (532 V/cm). After 48 h, <t>RNA</t> was extracted from skin biopsies. eGn mRNA levels were measured using <t>qRT-PCR,</t> normalized with RPS24 ribosomal RNA and expressed as 2 −ΔCT values (1 repeated experiment). Absence of residual plasmid in the RNA preparations was controlled using qPCR without reverse transcription
    Reverse Transcribed Rna, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 157 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Genecopoeia reverse transcribed cdna
    Design and expression of peGn, pscDEC-eGn and pscCD11c-eGn. a Schematic representation of peGn, pscDEC-eGn and pscCD11c-eGn. peGn includes the codon-optimized eGn sequence with its intrinsic signal peptide (SP, RVFV M segment from nt 411 to nt 1763) and cloned in the pcDNA4-V5His expression vector. pscDEC-eGn and pscCD11c include the signal peptide (SP) of the VH chain of the parental mAb sequence, the scFv (VH and VL) sequence and the nt 477 to nt 1763 portion of the codon-optimized eGn sequence. The VH and VL sequences are connected together with a (G 4 S) 4 linker and the scFv and eGn sequences are connected with a (G 4 S) 3 linker. These chimeric sequences were cloned in a pcDNA3.1 vector. b, c HEK293 cells were transfected with pcDNA3.1, peGn, pscDEC-eGn and pscCD11c-eGn. In ( b ), the cell lysates (10 µl) were separated by SDS-PAGE under reducing conditions and eGn was revealed with anti-RVFV HMAF followed by a HRP-GAM IgG. The predicted sizes of the expressed untargeted and chimeric eGn proteins are 48.8 kDa for untargeted eGn, 76.3 kDa for the scDEC-eGn protein and 78.6 kDa for the scCD11c-eGn protein. All samples were gathered from the same experiment and were processed in parallel (1 repeated experiment). In ( c ) the cell lysates (100 µl) and the concentrated cell supernatants (500 µl) gathered from the same experiment were spotted onto the same nitrocellulose membrane and the eGn protein was revealed as in ( b ) (1 repeated experiment). d Sheep skin lymph low-density cells were reacted with the parental anti-DEC205, anti-CD11c or isotype control mAbs (top panel) or with the concentrated supernatant from HEK293 cells transfected with peGn, pscDEC-eGn, pscCD11c-eGn and a negative control pcDNA3.1 (bottom panel). The staining of the DAPI neg FSC hi cells, which include dominantly cDCs, is depicted (dark gray) compared to control (dash line). Bound mAbs were revealed with A488-DAM IgG and bound eGn with anti-RVFV HMAF followed by A488-DAM IgG. e Relative expression of peGn, pscDEC-eGn and pscCD11c-eGn in sheep skin. peGn, pscDEC-eGn and pscCD11c-eGn (100 µg) were injected intradermally in 100 µl saline in inner front leg sites of two sheep followed by SEP (532 V/cm). After 48 h, <t>RNA</t> was extracted from skin biopsies. eGn mRNA levels were measured using <t>qRT-PCR,</t> normalized with RPS24 ribosomal RNA and expressed as 2 −ΔCT values (1 repeated experiment). Absence of residual plasmid in the RNA preparations was controlled using qPCR without reverse transcription
    Reverse Transcribed Cdna, supplied by Genecopoeia, used in various techniques. Bioz Stars score: 93/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Stratagene reverse transcribed cdna
    Design and expression of peGn, pscDEC-eGn and pscCD11c-eGn. a Schematic representation of peGn, pscDEC-eGn and pscCD11c-eGn. peGn includes the codon-optimized eGn sequence with its intrinsic signal peptide (SP, RVFV M segment from nt 411 to nt 1763) and cloned in the pcDNA4-V5His expression vector. pscDEC-eGn and pscCD11c include the signal peptide (SP) of the VH chain of the parental mAb sequence, the scFv (VH and VL) sequence and the nt 477 to nt 1763 portion of the codon-optimized eGn sequence. The VH and VL sequences are connected together with a (G 4 S) 4 linker and the scFv and eGn sequences are connected with a (G 4 S) 3 linker. These chimeric sequences were cloned in a pcDNA3.1 vector. b, c HEK293 cells were transfected with pcDNA3.1, peGn, pscDEC-eGn and pscCD11c-eGn. In ( b ), the cell lysates (10 µl) were separated by SDS-PAGE under reducing conditions and eGn was revealed with anti-RVFV HMAF followed by a HRP-GAM IgG. The predicted sizes of the expressed untargeted and chimeric eGn proteins are 48.8 kDa for untargeted eGn, 76.3 kDa for the scDEC-eGn protein and 78.6 kDa for the scCD11c-eGn protein. All samples were gathered from the same experiment and were processed in parallel (1 repeated experiment). In ( c ) the cell lysates (100 µl) and the concentrated cell supernatants (500 µl) gathered from the same experiment were spotted onto the same nitrocellulose membrane and the eGn protein was revealed as in ( b ) (1 repeated experiment). d Sheep skin lymph low-density cells were reacted with the parental anti-DEC205, anti-CD11c or isotype control mAbs (top panel) or with the concentrated supernatant from HEK293 cells transfected with peGn, pscDEC-eGn, pscCD11c-eGn and a negative control pcDNA3.1 (bottom panel). The staining of the DAPI neg FSC hi cells, which include dominantly cDCs, is depicted (dark gray) compared to control (dash line). Bound mAbs were revealed with A488-DAM IgG and bound eGn with anti-RVFV HMAF followed by A488-DAM IgG. e Relative expression of peGn, pscDEC-eGn and pscCD11c-eGn in sheep skin. peGn, pscDEC-eGn and pscCD11c-eGn (100 µg) were injected intradermally in 100 µl saline in inner front leg sites of two sheep followed by SEP (532 V/cm). After 48 h, <t>RNA</t> was extracted from skin biopsies. eGn mRNA levels were measured using <t>qRT-PCR,</t> normalized with RPS24 ribosomal RNA and expressed as 2 −ΔCT values (1 repeated experiment). Absence of residual plasmid in the RNA preparations was controlled using qPCR without reverse transcription
    Reverse Transcribed Cdna, supplied by Stratagene, used in various techniques. Bioz Stars score: 94/100, based on 62 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    TaKaRa reverse transcribed complementary dna
    Design and expression of peGn, pscDEC-eGn and pscCD11c-eGn. a Schematic representation of peGn, pscDEC-eGn and pscCD11c-eGn. peGn includes the codon-optimized eGn sequence with its intrinsic signal peptide (SP, RVFV M segment from nt 411 to nt 1763) and cloned in the pcDNA4-V5His expression vector. pscDEC-eGn and pscCD11c include the signal peptide (SP) of the VH chain of the parental mAb sequence, the scFv (VH and VL) sequence and the nt 477 to nt 1763 portion of the codon-optimized eGn sequence. The VH and VL sequences are connected together with a (G 4 S) 4 linker and the scFv and eGn sequences are connected with a (G 4 S) 3 linker. These chimeric sequences were cloned in a pcDNA3.1 vector. b, c HEK293 cells were transfected with pcDNA3.1, peGn, pscDEC-eGn and pscCD11c-eGn. In ( b ), the cell lysates (10 µl) were separated by SDS-PAGE under reducing conditions and eGn was revealed with anti-RVFV HMAF followed by a HRP-GAM IgG. The predicted sizes of the expressed untargeted and chimeric eGn proteins are 48.8 kDa for untargeted eGn, 76.3 kDa for the scDEC-eGn protein and 78.6 kDa for the scCD11c-eGn protein. All samples were gathered from the same experiment and were processed in parallel (1 repeated experiment). In ( c ) the cell lysates (100 µl) and the concentrated cell supernatants (500 µl) gathered from the same experiment were spotted onto the same nitrocellulose membrane and the eGn protein was revealed as in ( b ) (1 repeated experiment). d Sheep skin lymph low-density cells were reacted with the parental anti-DEC205, anti-CD11c or isotype control mAbs (top panel) or with the concentrated supernatant from HEK293 cells transfected with peGn, pscDEC-eGn, pscCD11c-eGn and a negative control pcDNA3.1 (bottom panel). The staining of the DAPI neg FSC hi cells, which include dominantly cDCs, is depicted (dark gray) compared to control (dash line). Bound mAbs were revealed with A488-DAM IgG and bound eGn with anti-RVFV HMAF followed by A488-DAM IgG. e Relative expression of peGn, pscDEC-eGn and pscCD11c-eGn in sheep skin. peGn, pscDEC-eGn and pscCD11c-eGn (100 µg) were injected intradermally in 100 µl saline in inner front leg sites of two sheep followed by SEP (532 V/cm). After 48 h, <t>RNA</t> was extracted from skin biopsies. eGn mRNA levels were measured using <t>qRT-PCR,</t> normalized with RPS24 ribosomal RNA and expressed as 2 −ΔCT values (1 repeated experiment). Absence of residual plasmid in the RNA preparations was controlled using qPCR without reverse transcription
    Reverse Transcribed Complementary Dna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 158 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Toyobo reverse transcribed cdna
    Tissue-specific distribution of transmembrane E3 ligases. Total RNAs from 23 human or 22 mouse tissues were reverse-transcribed and measured by <t>TaqMan-based</t> real-time PCR assay using the delta–delta Ct method. Data are normalised to the amount of 18S ribosomal RNA; results are expressed as the fold increase compared with <t>cDNA</t> pools from each tissue. ( a ) RNF183, ( b ) RNF186, ( c ) ZNRF4, ( d ) RNF182, ( e ) RNF150, ( f ) RNF175.
    Reverse Transcribed Cdna, supplied by Toyobo, used in various techniques. Bioz Stars score: 90/100, based on 82 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Bio-Rad reverse transcribed mrna
    Tissue-specific distribution of transmembrane E3 ligases. Total RNAs from 23 human or 22 mouse tissues were reverse-transcribed and measured by <t>TaqMan-based</t> real-time PCR assay using the delta–delta Ct method. Data are normalised to the amount of 18S ribosomal RNA; results are expressed as the fold increase compared with <t>cDNA</t> pools from each tissue. ( a ) RNF183, ( b ) RNF186, ( c ) ZNRF4, ( d ) RNF182, ( e ) RNF150, ( f ) RNF175.
    Reverse Transcribed Mrna, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 79/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Eurofins reverse transcribed rna
    Tissue-specific distribution of transmembrane E3 ligases. Total RNAs from 23 human or 22 mouse tissues were reverse-transcribed and measured by <t>TaqMan-based</t> real-time PCR assay using the delta–delta Ct method. Data are normalised to the amount of 18S ribosomal RNA; results are expressed as the fold increase compared with <t>cDNA</t> pools from each tissue. ( a ) RNF183, ( b ) RNF186, ( c ) ZNRF4, ( d ) RNF182, ( e ) RNF150, ( f ) RNF175.
    Reverse Transcribed Rna, supplied by Eurofins, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Design and expression of peGn, pscDEC-eGn and pscCD11c-eGn. a Schematic representation of peGn, pscDEC-eGn and pscCD11c-eGn. peGn includes the codon-optimized eGn sequence with its intrinsic signal peptide (SP, RVFV M segment from nt 411 to nt 1763) and cloned in the pcDNA4-V5His expression vector. pscDEC-eGn and pscCD11c include the signal peptide (SP) of the VH chain of the parental mAb sequence, the scFv (VH and VL) sequence and the nt 477 to nt 1763 portion of the codon-optimized eGn sequence. The VH and VL sequences are connected together with a (G 4 S) 4 linker and the scFv and eGn sequences are connected with a (G 4 S) 3 linker. These chimeric sequences were cloned in a pcDNA3.1 vector. b, c HEK293 cells were transfected with pcDNA3.1, peGn, pscDEC-eGn and pscCD11c-eGn. In ( b ), the cell lysates (10 µl) were separated by SDS-PAGE under reducing conditions and eGn was revealed with anti-RVFV HMAF followed by a HRP-GAM IgG. The predicted sizes of the expressed untargeted and chimeric eGn proteins are 48.8 kDa for untargeted eGn, 76.3 kDa for the scDEC-eGn protein and 78.6 kDa for the scCD11c-eGn protein. All samples were gathered from the same experiment and were processed in parallel (1 repeated experiment). In ( c ) the cell lysates (100 µl) and the concentrated cell supernatants (500 µl) gathered from the same experiment were spotted onto the same nitrocellulose membrane and the eGn protein was revealed as in ( b ) (1 repeated experiment). d Sheep skin lymph low-density cells were reacted with the parental anti-DEC205, anti-CD11c or isotype control mAbs (top panel) or with the concentrated supernatant from HEK293 cells transfected with peGn, pscDEC-eGn, pscCD11c-eGn and a negative control pcDNA3.1 (bottom panel). The staining of the DAPI neg FSC hi cells, which include dominantly cDCs, is depicted (dark gray) compared to control (dash line). Bound mAbs were revealed with A488-DAM IgG and bound eGn with anti-RVFV HMAF followed by A488-DAM IgG. e Relative expression of peGn, pscDEC-eGn and pscCD11c-eGn in sheep skin. peGn, pscDEC-eGn and pscCD11c-eGn (100 µg) were injected intradermally in 100 µl saline in inner front leg sites of two sheep followed by SEP (532 V/cm). After 48 h, RNA was extracted from skin biopsies. eGn mRNA levels were measured using qRT-PCR, normalized with RPS24 ribosomal RNA and expressed as 2 −ΔCT values (1 repeated experiment). Absence of residual plasmid in the RNA preparations was controlled using qPCR without reverse transcription

    Journal: NPJ Vaccines

    Article Title: A Rift Valley fever virus Gn ectodomain-based DNA vaccine induces a partial protection not improved by APC targeting

    doi: 10.1038/s41541-018-0052-x

    Figure Lengend Snippet: Design and expression of peGn, pscDEC-eGn and pscCD11c-eGn. a Schematic representation of peGn, pscDEC-eGn and pscCD11c-eGn. peGn includes the codon-optimized eGn sequence with its intrinsic signal peptide (SP, RVFV M segment from nt 411 to nt 1763) and cloned in the pcDNA4-V5His expression vector. pscDEC-eGn and pscCD11c include the signal peptide (SP) of the VH chain of the parental mAb sequence, the scFv (VH and VL) sequence and the nt 477 to nt 1763 portion of the codon-optimized eGn sequence. The VH and VL sequences are connected together with a (G 4 S) 4 linker and the scFv and eGn sequences are connected with a (G 4 S) 3 linker. These chimeric sequences were cloned in a pcDNA3.1 vector. b, c HEK293 cells were transfected with pcDNA3.1, peGn, pscDEC-eGn and pscCD11c-eGn. In ( b ), the cell lysates (10 µl) were separated by SDS-PAGE under reducing conditions and eGn was revealed with anti-RVFV HMAF followed by a HRP-GAM IgG. The predicted sizes of the expressed untargeted and chimeric eGn proteins are 48.8 kDa for untargeted eGn, 76.3 kDa for the scDEC-eGn protein and 78.6 kDa for the scCD11c-eGn protein. All samples were gathered from the same experiment and were processed in parallel (1 repeated experiment). In ( c ) the cell lysates (100 µl) and the concentrated cell supernatants (500 µl) gathered from the same experiment were spotted onto the same nitrocellulose membrane and the eGn protein was revealed as in ( b ) (1 repeated experiment). d Sheep skin lymph low-density cells were reacted with the parental anti-DEC205, anti-CD11c or isotype control mAbs (top panel) or with the concentrated supernatant from HEK293 cells transfected with peGn, pscDEC-eGn, pscCD11c-eGn and a negative control pcDNA3.1 (bottom panel). The staining of the DAPI neg FSC hi cells, which include dominantly cDCs, is depicted (dark gray) compared to control (dash line). Bound mAbs were revealed with A488-DAM IgG and bound eGn with anti-RVFV HMAF followed by A488-DAM IgG. e Relative expression of peGn, pscDEC-eGn and pscCD11c-eGn in sheep skin. peGn, pscDEC-eGn and pscCD11c-eGn (100 µg) were injected intradermally in 100 µl saline in inner front leg sites of two sheep followed by SEP (532 V/cm). After 48 h, RNA was extracted from skin biopsies. eGn mRNA levels were measured using qRT-PCR, normalized with RPS24 ribosomal RNA and expressed as 2 −ΔCT values (1 repeated experiment). Absence of residual plasmid in the RNA preparations was controlled using qPCR without reverse transcription

    Article Snippet: In order to check that the transfected skin RNA did not include detectable residual plasmids, reverse-transcribed (3 ng estimated cDNA) and non-reverse transcribed RNA (3 ng RNA) were subjected to PCR reaction with the iTaq UniverSYBR Green (Bio-Rad) for amplifying a Gn fragment with forward primer 5′ – AGT GCG ATG GGC AGT TGT C–3′ (M segment, codon-optimized sequence) and reverse primer 5′ – TTC TTG AAC ACG GCA AAT GG–3′.

    Techniques: Expressing, Sequencing, Clone Assay, Plasmid Preparation, Transfection, SDS Page, Negative Control, Staining, Injection, Quantitative RT-PCR, Real-time Polymerase Chain Reaction

    Tissue-specific distribution of transmembrane E3 ligases. Total RNAs from 23 human or 22 mouse tissues were reverse-transcribed and measured by TaqMan-based real-time PCR assay using the delta–delta Ct method. Data are normalised to the amount of 18S ribosomal RNA; results are expressed as the fold increase compared with cDNA pools from each tissue. ( a ) RNF183, ( b ) RNF186, ( c ) ZNRF4, ( d ) RNF182, ( e ) RNF150, ( f ) RNF175.

    Journal: Scientific Reports

    Article Title: Genome-wide identification and gene expression profiling of ubiquitin ligases for endoplasmic reticulum protein degradation

    doi: 10.1038/srep30955

    Figure Lengend Snippet: Tissue-specific distribution of transmembrane E3 ligases. Total RNAs from 23 human or 22 mouse tissues were reverse-transcribed and measured by TaqMan-based real-time PCR assay using the delta–delta Ct method. Data are normalised to the amount of 18S ribosomal RNA; results are expressed as the fold increase compared with cDNA pools from each tissue. ( a ) RNF183, ( b ) RNF186, ( c ) ZNRF4, ( d ) RNF182, ( e ) RNF150, ( f ) RNF175.

    Article Snippet: The reverse-transcribed cDNA was measured by TaqMan-based real-time PCR assay using the THUNDERBIRD Probe qPCR Mix (TOYOBO) on a LightCycler 480 Instrument II real-time PCR system (Roche diagnostics, Basel, Switzerland).

    Techniques: Real-time Polymerase Chain Reaction