tandem-dimer red fluorescent protein reporter Search Results


ms2 bacteriophage coat protein tandem dimer  (Addgene inc)
95
Addgene inc ms2 bacteriophage coat protein tandem dimer
Evaluation of RNA loading into EVs via the TAMEL platform. (a) This cartoon summarizes the concept of facilitating active loading of cargo RNA into EVs via our TAMEL platform. A TAMEL EV-loading protein comprises an EV-enriched protein (EEP, blue) fused to an RNA-binding domain (RBD, green), which localizes to EVs. Actively loaded RNA (green) contains a motif that binds to the RBD, resulting in enhanced loading into EVs relative to passively loaded RNA (orange). (b) RNA cargo design impacts active loading. The “fold enrichment of cargo mRNA <t>+/−MS2”</t> is defined as the ratio of cargo RNA/GAPDH mRNA in EVs derived from cells expressing Lamp2b–MS2–HA divided by the same RNA ratio in EVs derived from cells expressing Lamp2b–HA. All experiments were performed in biological triplicates. (c) Cartoon illustrating the 3′ RNA fragment analysis technique. Cargo RNA is first reverse transcribed using an oligo dT primer, and amplicons corresponding to the RNA 5′ or 3′ ends (the latter is located ~500 bases upstream of the polyA site) are then quantified by qPCR using the primer pairs indicated. Note that the amplicon near the RNA 3′ end will be present in cDNA derived from both full-length RNA and 3′ RNA fragments. (d) Analysis of 3′ RNA fragment loading into EVs. Cargo RNA levels were quantified as depicted in panel c and normalized to GAPDH. Passive loading: cells transfected with Lamp2b–HA; active loading: cells transfected with Lamp2b–MS2–HA. (e) Full-length RNA and 3′ fragment RNA levels in EVs were quantified following incubation at 37°C; experiments were performed in technical duplicate with a biological replicate shown in Supplementary Fig. 2a. Error bars indicate 1 standard deviation, throughout. MVB, multivesicular body.
Ms2 Bacteriophage Coat Protein Tandem Dimer, supplied by Addgene inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/tandem-dimer+red+fluorescent+protein+reporter/pMS2-GFP+(Plasmid+%2327121)/pmc04870355-22-6-22
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peb3 tdtomato  (Addgene inc)
93
Addgene inc peb3 tdtomato
Evaluation of RNA loading into EVs via the TAMEL platform. (a) This cartoon summarizes the concept of facilitating active loading of cargo RNA into EVs via our TAMEL platform. A TAMEL EV-loading protein comprises an EV-enriched protein (EEP, blue) fused to an RNA-binding domain (RBD, green), which localizes to EVs. Actively loaded RNA (green) contains a motif that binds to the RBD, resulting in enhanced loading into EVs relative to passively loaded RNA (orange). (b) RNA cargo design impacts active loading. The “fold enrichment of cargo mRNA <t>+/−MS2”</t> is defined as the ratio of cargo RNA/GAPDH mRNA in EVs derived from cells expressing Lamp2b–MS2–HA divided by the same RNA ratio in EVs derived from cells expressing Lamp2b–HA. All experiments were performed in biological triplicates. (c) Cartoon illustrating the 3′ RNA fragment analysis technique. Cargo RNA is first reverse transcribed using an oligo dT primer, and amplicons corresponding to the RNA 5′ or 3′ ends (the latter is located ~500 bases upstream of the polyA site) are then quantified by qPCR using the primer pairs indicated. Note that the amplicon near the RNA 3′ end will be present in cDNA derived from both full-length RNA and 3′ RNA fragments. (d) Analysis of 3′ RNA fragment loading into EVs. Cargo RNA levels were quantified as depicted in panel c and normalized to GAPDH. Passive loading: cells transfected with Lamp2b–HA; active loading: cells transfected with Lamp2b–MS2–HA. (e) Full-length RNA and 3′ fragment RNA levels in EVs were quantified following incubation at 37°C; experiments were performed in technical duplicate with a biological replicate shown in Supplementary Fig. 2a. Error bars indicate 1 standard deviation, throughout. MVB, multivesicular body.
Peb3 Tdtomato, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/tandem-dimer+red+fluorescent+protein+reporter/EB3-tdTomato+(Plasmid+%2350708)/pmc07564071-227-4-20
Average 93 stars, based on 1 article reviews
peb3 tdtomato - by Bioz Stars, 2026-09
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tandem dimer addgene  (Addgene inc)
90
Addgene inc tandem dimer addgene
Evaluation of RNA loading into EVs via the TAMEL platform. (a) This cartoon summarizes the concept of facilitating active loading of cargo RNA into EVs via our TAMEL platform. A TAMEL EV-loading protein comprises an EV-enriched protein (EEP, blue) fused to an RNA-binding domain (RBD, green), which localizes to EVs. Actively loaded RNA (green) contains a motif that binds to the RBD, resulting in enhanced loading into EVs relative to passively loaded RNA (orange). (b) RNA cargo design impacts active loading. The “fold enrichment of cargo mRNA <t>+/−MS2”</t> is defined as the ratio of cargo RNA/GAPDH mRNA in EVs derived from cells expressing Lamp2b–MS2–HA divided by the same RNA ratio in EVs derived from cells expressing Lamp2b–HA. All experiments were performed in biological triplicates. (c) Cartoon illustrating the 3′ RNA fragment analysis technique. Cargo RNA is first reverse transcribed using an oligo dT primer, and amplicons corresponding to the RNA 5′ or 3′ ends (the latter is located ~500 bases upstream of the polyA site) are then quantified by qPCR using the primer pairs indicated. Note that the amplicon near the RNA 3′ end will be present in cDNA derived from both full-length RNA and 3′ RNA fragments. (d) Analysis of 3′ RNA fragment loading into EVs. Cargo RNA levels were quantified as depicted in panel c and normalized to GAPDH. Passive loading: cells transfected with Lamp2b–HA; active loading: cells transfected with Lamp2b–MS2–HA. (e) Full-length RNA and 3′ fragment RNA levels in EVs were quantified following incubation at 37°C; experiments were performed in technical duplicate with a biological replicate shown in Supplementary Fig. 2a. Error bars indicate 1 standard deviation, throughout. MVB, multivesicular body.
Tandem Dimer Addgene, supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/tandem-dimer+red+fluorescent+protein+reporter/pCFJ1955+(Plasmid+%2384830)/pm35766819-103-101-103
Average 90 stars, based on 1 article reviews
tandem dimer addgene - by Bioz Stars, 2026-09
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pspcas9 bb 2a gfp px458  (Addgene inc)
96
Addgene inc pspcas9 bb 2a gfp px458
Evaluation of RNA loading into EVs via the TAMEL platform. (a) This cartoon summarizes the concept of facilitating active loading of cargo RNA into EVs via our TAMEL platform. A TAMEL EV-loading protein comprises an EV-enriched protein (EEP, blue) fused to an RNA-binding domain (RBD, green), which localizes to EVs. Actively loaded RNA (green) contains a motif that binds to the RBD, resulting in enhanced loading into EVs relative to passively loaded RNA (orange). (b) RNA cargo design impacts active loading. The “fold enrichment of cargo mRNA <t>+/−MS2”</t> is defined as the ratio of cargo RNA/GAPDH mRNA in EVs derived from cells expressing Lamp2b–MS2–HA divided by the same RNA ratio in EVs derived from cells expressing Lamp2b–HA. All experiments were performed in biological triplicates. (c) Cartoon illustrating the 3′ RNA fragment analysis technique. Cargo RNA is first reverse transcribed using an oligo dT primer, and amplicons corresponding to the RNA 5′ or 3′ ends (the latter is located ~500 bases upstream of the polyA site) are then quantified by qPCR using the primer pairs indicated. Note that the amplicon near the RNA 3′ end will be present in cDNA derived from both full-length RNA and 3′ RNA fragments. (d) Analysis of 3′ RNA fragment loading into EVs. Cargo RNA levels were quantified as depicted in panel c and normalized to GAPDH. Passive loading: cells transfected with Lamp2b–HA; active loading: cells transfected with Lamp2b–MS2–HA. (e) Full-length RNA and 3′ fragment RNA levels in EVs were quantified following incubation at 37°C; experiments were performed in technical duplicate with a biological replicate shown in Supplementary Fig. 2a. Error bars indicate 1 standard deviation, throughout. MVB, multivesicular body.
Pspcas9 Bb 2a Gfp Px458, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/tandem-dimer+red+fluorescent+protein+reporter/pSpCas9(BB)-2A-GFP+(PX458)+(Plasmid+%2348138)/pm29981367-350-22-30
Average 96 stars, based on 1 article reviews
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rosa26 locus  (Jackson Laboratory)
86
Jackson Laboratory rosa26 locus
Evaluation of RNA loading into EVs via the TAMEL platform. (a) This cartoon summarizes the concept of facilitating active loading of cargo RNA into EVs via our TAMEL platform. A TAMEL EV-loading protein comprises an EV-enriched protein (EEP, blue) fused to an RNA-binding domain (RBD, green), which localizes to EVs. Actively loaded RNA (green) contains a motif that binds to the RBD, resulting in enhanced loading into EVs relative to passively loaded RNA (orange). (b) RNA cargo design impacts active loading. The “fold enrichment of cargo mRNA <t>+/−MS2”</t> is defined as the ratio of cargo RNA/GAPDH mRNA in EVs derived from cells expressing Lamp2b–MS2–HA divided by the same RNA ratio in EVs derived from cells expressing Lamp2b–HA. All experiments were performed in biological triplicates. (c) Cartoon illustrating the 3′ RNA fragment analysis technique. Cargo RNA is first reverse transcribed using an oligo dT primer, and amplicons corresponding to the RNA 5′ or 3′ ends (the latter is located ~500 bases upstream of the polyA site) are then quantified by qPCR using the primer pairs indicated. Note that the amplicon near the RNA 3′ end will be present in cDNA derived from both full-length RNA and 3′ RNA fragments. (d) Analysis of 3′ RNA fragment loading into EVs. Cargo RNA levels were quantified as depicted in panel c and normalized to GAPDH. Passive loading: cells transfected with Lamp2b–HA; active loading: cells transfected with Lamp2b–MS2–HA. (e) Full-length RNA and 3′ fragment RNA levels in EVs were quantified following incubation at 37°C; experiments were performed in technical duplicate with a biological replicate shown in Supplementary Fig. 2a. Error bars indicate 1 standard deviation, throughout. MVB, multivesicular body.
Rosa26 Locus, supplied by Jackson Laboratory, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/tandem-dimer+red+fluorescent+protein+reporter/locus+rosa26/pmc12883771-100-27-36
Average 86 stars, based on 1 article reviews
rosa26 locus - by Bioz Stars, 2026-09
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ef1 tdtomato lentiviral vector  (Vectalys Inc)
90
Vectalys Inc ef1 tdtomato lentiviral vector
Evaluation of RNA loading into EVs via the TAMEL platform. (a) This cartoon summarizes the concept of facilitating active loading of cargo RNA into EVs via our TAMEL platform. A TAMEL EV-loading protein comprises an EV-enriched protein (EEP, blue) fused to an RNA-binding domain (RBD, green), which localizes to EVs. Actively loaded RNA (green) contains a motif that binds to the RBD, resulting in enhanced loading into EVs relative to passively loaded RNA (orange). (b) RNA cargo design impacts active loading. The “fold enrichment of cargo mRNA <t>+/−MS2”</t> is defined as the ratio of cargo RNA/GAPDH mRNA in EVs derived from cells expressing Lamp2b–MS2–HA divided by the same RNA ratio in EVs derived from cells expressing Lamp2b–HA. All experiments were performed in biological triplicates. (c) Cartoon illustrating the 3′ RNA fragment analysis technique. Cargo RNA is first reverse transcribed using an oligo dT primer, and amplicons corresponding to the RNA 5′ or 3′ ends (the latter is located ~500 bases upstream of the polyA site) are then quantified by qPCR using the primer pairs indicated. Note that the amplicon near the RNA 3′ end will be present in cDNA derived from both full-length RNA and 3′ RNA fragments. (d) Analysis of 3′ RNA fragment loading into EVs. Cargo RNA levels were quantified as depicted in panel c and normalized to GAPDH. Passive loading: cells transfected with Lamp2b–HA; active loading: cells transfected with Lamp2b–MS2–HA. (e) Full-length RNA and 3′ fragment RNA levels in EVs were quantified following incubation at 37°C; experiments were performed in technical duplicate with a biological replicate shown in Supplementary Fig. 2a. Error bars indicate 1 standard deviation, throughout. MVB, multivesicular body.
Ef1 Tdtomato Lentiviral Vector, supplied by Vectalys Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/tandem-dimer+red+fluorescent+protein+reporter/ef1+tdtomato+lentivector/pmc10665670-198-35-39
Average 90 stars, based on 1 article reviews
ef1 tdtomato lentiviral vector - by Bioz Stars, 2026-09
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epifluorescence stereomicroscope  (Olympus)
96
Olympus epifluorescence stereomicroscope
Evaluation of RNA loading into EVs via the TAMEL platform. (a) This cartoon summarizes the concept of facilitating active loading of cargo RNA into EVs via our TAMEL platform. A TAMEL EV-loading protein comprises an EV-enriched protein (EEP, blue) fused to an RNA-binding domain (RBD, green), which localizes to EVs. Actively loaded RNA (green) contains a motif that binds to the RBD, resulting in enhanced loading into EVs relative to passively loaded RNA (orange). (b) RNA cargo design impacts active loading. The “fold enrichment of cargo mRNA <t>+/−MS2”</t> is defined as the ratio of cargo RNA/GAPDH mRNA in EVs derived from cells expressing Lamp2b–MS2–HA divided by the same RNA ratio in EVs derived from cells expressing Lamp2b–HA. All experiments were performed in biological triplicates. (c) Cartoon illustrating the 3′ RNA fragment analysis technique. Cargo RNA is first reverse transcribed using an oligo dT primer, and amplicons corresponding to the RNA 5′ or 3′ ends (the latter is located ~500 bases upstream of the polyA site) are then quantified by qPCR using the primer pairs indicated. Note that the amplicon near the RNA 3′ end will be present in cDNA derived from both full-length RNA and 3′ RNA fragments. (d) Analysis of 3′ RNA fragment loading into EVs. Cargo RNA levels were quantified as depicted in panel c and normalized to GAPDH. Passive loading: cells transfected with Lamp2b–HA; active loading: cells transfected with Lamp2b–MS2–HA. (e) Full-length RNA and 3′ fragment RNA levels in EVs were quantified following incubation at 37°C; experiments were performed in technical duplicate with a biological replicate shown in Supplementary Fig. 2a. Error bars indicate 1 standard deviation, throughout. MVB, multivesicular body.
Epifluorescence Stereomicroscope, supplied by Olympus, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/tandem-dimer+red+fluorescent+protein+reporter/SZX7+Stereomicroscope+System/pmc05059792-392-8-10
Average 96 stars, based on 1 article reviews
epifluorescence stereomicroscope - by Bioz Stars, 2026-09
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ai9 reporter mice  (Jackson Laboratory)
86
Jackson Laboratory ai9 reporter mice
Evaluation of RNA loading into EVs via the TAMEL platform. (a) This cartoon summarizes the concept of facilitating active loading of cargo RNA into EVs via our TAMEL platform. A TAMEL EV-loading protein comprises an EV-enriched protein (EEP, blue) fused to an RNA-binding domain (RBD, green), which localizes to EVs. Actively loaded RNA (green) contains a motif that binds to the RBD, resulting in enhanced loading into EVs relative to passively loaded RNA (orange). (b) RNA cargo design impacts active loading. The “fold enrichment of cargo mRNA <t>+/−MS2”</t> is defined as the ratio of cargo RNA/GAPDH mRNA in EVs derived from cells expressing Lamp2b–MS2–HA divided by the same RNA ratio in EVs derived from cells expressing Lamp2b–HA. All experiments were performed in biological triplicates. (c) Cartoon illustrating the 3′ RNA fragment analysis technique. Cargo RNA is first reverse transcribed using an oligo dT primer, and amplicons corresponding to the RNA 5′ or 3′ ends (the latter is located ~500 bases upstream of the polyA site) are then quantified by qPCR using the primer pairs indicated. Note that the amplicon near the RNA 3′ end will be present in cDNA derived from both full-length RNA and 3′ RNA fragments. (d) Analysis of 3′ RNA fragment loading into EVs. Cargo RNA levels were quantified as depicted in panel c and normalized to GAPDH. Passive loading: cells transfected with Lamp2b–HA; active loading: cells transfected with Lamp2b–MS2–HA. (e) Full-length RNA and 3′ fragment RNA levels in EVs were quantified following incubation at 37°C; experiments were performed in technical duplicate with a biological replicate shown in Supplementary Fig. 2a. Error bars indicate 1 standard deviation, throughout. MVB, multivesicular body.
Ai9 Reporter Mice, supplied by Jackson Laboratory, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/tandem-dimer+red+fluorescent+protein+reporter/ai9/pm41348674-55-19-22
Average 86 stars, based on 1 article reviews
ai9 reporter mice - by Bioz Stars, 2026-09
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gfp tdmcp gfp  (Addgene inc)
93
Addgene inc gfp tdmcp gfp
Overview of recombinant baculoviruses used in this study. All recombinant proteins were expressed with Myc-His tag at the C-terminus.
Gfp Tdmcp Gfp, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/tandem-dimer+red+fluorescent+protein+reporter/phage-ubc-nls-ha-tdMCP-gfp+(Plasmid+%2340649)/pmc10926400-72-32-40
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gfp mapper  (Addgene inc)
93
Addgene inc gfp mapper
Overview of recombinant baculoviruses used in this study. All recombinant proteins were expressed with Myc-His tag at the C-terminus.
Gfp Mapper, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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helios gene gun  (Bio-Rad)
96
Bio-Rad helios gene gun
Overview of recombinant baculoviruses used in this study. All recombinant proteins were expressed with Myc-His tag at the C-terminus.
Helios Gene Gun, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/tandem-dimer+red+fluorescent+protein+reporter/Helios+Gene+Gun+System/pmc04883118-535-51-54
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edward boyden  (Addgene inc)
97
Addgene inc edward boyden
Overview of recombinant baculoviruses used in this study. All recombinant proteins were expressed with Myc-His tag at the C-terminus.
Edward Boyden, supplied by Addgene inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/tandem-dimer+red+fluorescent+protein+reporter/pAAV-CAG-GFP+(Plasmid+%2337825)/pm36179767-67-22-24
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Image Search Results


Evaluation of RNA loading into EVs via the TAMEL platform. (a) This cartoon summarizes the concept of facilitating active loading of cargo RNA into EVs via our TAMEL platform. A TAMEL EV-loading protein comprises an EV-enriched protein (EEP, blue) fused to an RNA-binding domain (RBD, green), which localizes to EVs. Actively loaded RNA (green) contains a motif that binds to the RBD, resulting in enhanced loading into EVs relative to passively loaded RNA (orange). (b) RNA cargo design impacts active loading. The “fold enrichment of cargo mRNA +/−MS2” is defined as the ratio of cargo RNA/GAPDH mRNA in EVs derived from cells expressing Lamp2b–MS2–HA divided by the same RNA ratio in EVs derived from cells expressing Lamp2b–HA. All experiments were performed in biological triplicates. (c) Cartoon illustrating the 3′ RNA fragment analysis technique. Cargo RNA is first reverse transcribed using an oligo dT primer, and amplicons corresponding to the RNA 5′ or 3′ ends (the latter is located ~500 bases upstream of the polyA site) are then quantified by qPCR using the primer pairs indicated. Note that the amplicon near the RNA 3′ end will be present in cDNA derived from both full-length RNA and 3′ RNA fragments. (d) Analysis of 3′ RNA fragment loading into EVs. Cargo RNA levels were quantified as depicted in panel c and normalized to GAPDH. Passive loading: cells transfected with Lamp2b–HA; active loading: cells transfected with Lamp2b–MS2–HA. (e) Full-length RNA and 3′ fragment RNA levels in EVs were quantified following incubation at 37°C; experiments were performed in technical duplicate with a biological replicate shown in Supplementary Fig. 2a. Error bars indicate 1 standard deviation, throughout. MVB, multivesicular body.

Journal: Journal of Extracellular Vesicles

Article Title: A platform for actively loading cargo RNA to elucidate limiting steps in EV-mediated delivery

doi: 10.3402/jev.v5.31027

Figure Lengend Snippet: Evaluation of RNA loading into EVs via the TAMEL platform. (a) This cartoon summarizes the concept of facilitating active loading of cargo RNA into EVs via our TAMEL platform. A TAMEL EV-loading protein comprises an EV-enriched protein (EEP, blue) fused to an RNA-binding domain (RBD, green), which localizes to EVs. Actively loaded RNA (green) contains a motif that binds to the RBD, resulting in enhanced loading into EVs relative to passively loaded RNA (orange). (b) RNA cargo design impacts active loading. The “fold enrichment of cargo mRNA +/−MS2” is defined as the ratio of cargo RNA/GAPDH mRNA in EVs derived from cells expressing Lamp2b–MS2–HA divided by the same RNA ratio in EVs derived from cells expressing Lamp2b–HA. All experiments were performed in biological triplicates. (c) Cartoon illustrating the 3′ RNA fragment analysis technique. Cargo RNA is first reverse transcribed using an oligo dT primer, and amplicons corresponding to the RNA 5′ or 3′ ends (the latter is located ~500 bases upstream of the polyA site) are then quantified by qPCR using the primer pairs indicated. Note that the amplicon near the RNA 3′ end will be present in cDNA derived from both full-length RNA and 3′ RNA fragments. (d) Analysis of 3′ RNA fragment loading into EVs. Cargo RNA levels were quantified as depicted in panel c and normalized to GAPDH. Passive loading: cells transfected with Lamp2b–HA; active loading: cells transfected with Lamp2b–MS2–HA. (e) Full-length RNA and 3′ fragment RNA levels in EVs were quantified following incubation at 37°C; experiments were performed in technical duplicate with a biological replicate shown in Supplementary Fig. 2a. Error bars indicate 1 standard deviation, throughout. MVB, multivesicular body.

Article Snippet: The pMS2-GFP plasmid, which encodes the MS2 bacteriophage coat protein tandem dimer (mutant d1FG and V29I ( , )), was obtained from Addgene (plasmid #27121), deposited by Robert Singer ( ).

Techniques: RNA Binding Assay, Derivative Assay, Expressing, Reverse Transcription, Amplification, Transfection, Incubation, Standard Deviation

Impact of EEP choice on TAMEL-mediated active RNA loading into vesicles. (a) Effects of EEP choice on cargo RNA loading into EVs. Experiments were performed in biological triplicate. (b) Effects of EEP choice on cargo loading into gesicles. Experiments were performed in biological triplicate. (c) Protein abundance was quantified by densitometry analysis of anti-HA western blots (Supplementary Fig. 3), and each blot was internally normalized by the intensity for VSVG–MS2–HA in gesicles (maximal intensity case). This experiment was performed in biological duplicate. The y-axis is in log scale to enable visualization of all values. Error bars indicate 1 standard deviation, throughout.

Journal: Journal of Extracellular Vesicles

Article Title: A platform for actively loading cargo RNA to elucidate limiting steps in EV-mediated delivery

doi: 10.3402/jev.v5.31027

Figure Lengend Snippet: Impact of EEP choice on TAMEL-mediated active RNA loading into vesicles. (a) Effects of EEP choice on cargo RNA loading into EVs. Experiments were performed in biological triplicate. (b) Effects of EEP choice on cargo loading into gesicles. Experiments were performed in biological triplicate. (c) Protein abundance was quantified by densitometry analysis of anti-HA western blots (Supplementary Fig. 3), and each blot was internally normalized by the intensity for VSVG–MS2–HA in gesicles (maximal intensity case). This experiment was performed in biological duplicate. The y-axis is in log scale to enable visualization of all values. Error bars indicate 1 standard deviation, throughout.

Article Snippet: The pMS2-GFP plasmid, which encodes the MS2 bacteriophage coat protein tandem dimer (mutant d1FG and V29I ( , )), was obtained from Addgene (plasmid #27121), deposited by Robert Singer ( ).

Techniques: Quantitative Proteomics, Western Blot, Standard Deviation

Comparative analysis of dTomato delivery by actively or passively loaded vesicles. For panels a–c, the cartoons at left summarize the experimental designs, and in the panels at right, each data point represents the average of duplicate wells of cells treated with the same type of vesicle. Error bars indicate 1 standard deviation. “Normalized fluorescence” is defined as the mean fluorescence of cells receiving vesicles divided by the mean fluorescence of cells receiving a medium change only. (a) Time course of EV delivery to cells. Grey squares: CD63–HA EVs; orange circles: CD63–MS2–HA EVs. The solid arrow represents cells that received a medium change after 4 h of EV treatment. The dashed arrow indicates that cells did not receive a medium change. Statistically significant differences (p<0.05, not shown for clarity): CD63–MS2–HA +/− medium change. An independent repeat of this experiment is shown in Supplementary Fig. 3a. (b) Time course of gesicle delivery to cells. Purple squares: VSVG–HA gesicles; green circles: VSVG–MS2–HA gesicles. Solid and dashed arrows carry the same meaning as in panel (a). Statistically significant differences (p < 0.05, not shown for clarity): VSVG–HA versus VSVG–MS2–HA at 4 and 16 h (comparisons were made for each time point), VSVG–HA +/− medium change, and VSVG–MS2–HA +/− medium change. An independent repeat of this experiment is shown in Supplementary Fig. 3c. (c) Comparison of delivery by gesicles from cells transfected with VSVG–HA (purple), VSVG–MS2–HA (green) or a 50:50 mix of VSVG–HA and VSVG–MS2–HA (hybrid gesicles, magenta). (d) dTomato RNA levels (normalized to GAPDH) in VSVG–HA gesicles (purple), VSVG–MS2–HA gesicles (green) or hybrid gesicles (magenta). Error bars indicate 1 standard deviation of technical duplicate samples. *Significant difference was evaluated with a Student's t -test using a cut-off of p < 0.05.

Journal: Journal of Extracellular Vesicles

Article Title: A platform for actively loading cargo RNA to elucidate limiting steps in EV-mediated delivery

doi: 10.3402/jev.v5.31027

Figure Lengend Snippet: Comparative analysis of dTomato delivery by actively or passively loaded vesicles. For panels a–c, the cartoons at left summarize the experimental designs, and in the panels at right, each data point represents the average of duplicate wells of cells treated with the same type of vesicle. Error bars indicate 1 standard deviation. “Normalized fluorescence” is defined as the mean fluorescence of cells receiving vesicles divided by the mean fluorescence of cells receiving a medium change only. (a) Time course of EV delivery to cells. Grey squares: CD63–HA EVs; orange circles: CD63–MS2–HA EVs. The solid arrow represents cells that received a medium change after 4 h of EV treatment. The dashed arrow indicates that cells did not receive a medium change. Statistically significant differences (p<0.05, not shown for clarity): CD63–MS2–HA +/− medium change. An independent repeat of this experiment is shown in Supplementary Fig. 3a. (b) Time course of gesicle delivery to cells. Purple squares: VSVG–HA gesicles; green circles: VSVG–MS2–HA gesicles. Solid and dashed arrows carry the same meaning as in panel (a). Statistically significant differences (p < 0.05, not shown for clarity): VSVG–HA versus VSVG–MS2–HA at 4 and 16 h (comparisons were made for each time point), VSVG–HA +/− medium change, and VSVG–MS2–HA +/− medium change. An independent repeat of this experiment is shown in Supplementary Fig. 3c. (c) Comparison of delivery by gesicles from cells transfected with VSVG–HA (purple), VSVG–MS2–HA (green) or a 50:50 mix of VSVG–HA and VSVG–MS2–HA (hybrid gesicles, magenta). (d) dTomato RNA levels (normalized to GAPDH) in VSVG–HA gesicles (purple), VSVG–MS2–HA gesicles (green) or hybrid gesicles (magenta). Error bars indicate 1 standard deviation of technical duplicate samples. *Significant difference was evaluated with a Student's t -test using a cut-off of p < 0.05.

Article Snippet: The pMS2-GFP plasmid, which encodes the MS2 bacteriophage coat protein tandem dimer (mutant d1FG and V29I ( , )), was obtained from Addgene (plasmid #27121), deposited by Robert Singer ( ).

Techniques: Standard Deviation, Fluorescence, Comparison, Transfection

Overview of recombinant baculoviruses used in this study. All recombinant proteins were expressed with Myc-His tag at the C-terminus.

Journal: Frontiers in Insect Science

Article Title: Analysis of luciferase dsRNA production during baculovirus infection of Hi5 cells: RNA hairpins expressed by very late promoters do not trigger gene silencing

doi: 10.3389/finsc.2022.959077

Figure Lengend Snippet: Overview of recombinant baculoviruses used in this study. All recombinant proteins were expressed with Myc-His tag at the C-terminus.

Article Snippet: DsRNA-binding proteins such as the B2 RNAi suppressor protein of Flock house virus (FHV) , the fluorescent dsRNA-binding protein B2-GFP ( ) and the tandem dimer of MS2 coat protein fused to GFP (tdMCP-GFP) (amplified from plasmid phage-ubc-nls-ha-tdMCP-gfp; obtained from Addgene; ) were expressed by the p10 promoter as engineered in the pFastBac-Dual vector.

Techniques: Recombinant, Plasmid Preparation