mrna Thermo Fisher Search Results


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  • 94
    Thermo Fisher poly a tail length assay
    Transfection of non-phagocytic cells, using <t>mRNA:LPNs</t> and mRNA:CS-PLGA at different ratios performed in epithelial A549 cells for 24 h and 48 h post-transfection using flow cytometer. mRNA complexed LPNs reveal a significant higher transgene expression over CS-PLGA NPs. Both NPs show higher transfection rates with higher mRNA:NP ratios and increasing transfection until 48 h post-transfection N = 4, mean ± SD (*p
    Poly A Tail Length Assay, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 110 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore mrna
    UGT2B17 <t>mRNA</t> is transcribed in minor H antigen positive but not minor H antigen negative cells. (A) Northern blot analysis of total <t>RNA</t> from recipient and donor B-LCL, and from HLA-A29 + B-LCLs from 4 unrelated donors (designated 1–4). The blot was hybridized with a 32 P-labeled UGT2B17 probe for 16 h at 68°C. Detection of GAPDH mRNA was performed as a control. (B) Cytotoxicity assay for B-LCL by PL8 CTL. The B-LCL from unrelated donors ( 1 – 4 ) are the same lines used for analysis of gene expression in Fig. 3 A. Specific lysis is shown as the mean of triplicate cultures at an E:T ratio of 5:1. (C) Transfection of UGT2B17 cDNA into donor B-LCL restores recognition by PL8 CTL. Donor B-LCL were transfected by electroporation with the full-length construct of UGT2B17 , selected for 3 d with puromycin (0.6 μg/ml), and assayed as targets for PL8 CTL. The lysis of UGT2B17 -transfected donor B-LCL (triangles), untransfected donor B-LCL (circles), and recipient B-LCL (squares) is shown at various E:T ratios.
    Mrna, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2985 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Thermo Fisher poly a tail length assay kit
    UGT2B17 <t>mRNA</t> is transcribed in minor H antigen positive but not minor H antigen negative cells. (A) Northern blot analysis of total <t>RNA</t> from recipient and donor B-LCL, and from HLA-A29 + B-LCLs from 4 unrelated donors (designated 1–4). The blot was hybridized with a 32 P-labeled UGT2B17 probe for 16 h at 68°C. Detection of GAPDH mRNA was performed as a control. (B) Cytotoxicity assay for B-LCL by PL8 CTL. The B-LCL from unrelated donors ( 1 – 4 ) are the same lines used for analysis of gene expression in Fig. 3 A. Specific lysis is shown as the mean of triplicate cultures at an E:T ratio of 5:1. (C) Transfection of UGT2B17 cDNA into donor B-LCL restores recognition by PL8 CTL. Donor B-LCL were transfected by electroporation with the full-length construct of UGT2B17 , selected for 3 d with puromycin (0.6 μg/ml), and assayed as targets for PL8 CTL. The lysis of UGT2B17 -transfected donor B-LCL (triangles), untransfected donor B-LCL (circles), and recipient B-LCL (squares) is shown at various E:T ratios.
    Poly A Tail Length Assay Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher human stat3 mrna
    UGT2B17 <t>mRNA</t> is transcribed in minor H antigen positive but not minor H antigen negative cells. (A) Northern blot analysis of total <t>RNA</t> from recipient and donor B-LCL, and from HLA-A29 + B-LCLs from 4 unrelated donors (designated 1–4). The blot was hybridized with a 32 P-labeled UGT2B17 probe for 16 h at 68°C. Detection of GAPDH mRNA was performed as a control. (B) Cytotoxicity assay for B-LCL by PL8 CTL. The B-LCL from unrelated donors ( 1 – 4 ) are the same lines used for analysis of gene expression in Fig. 3 A. Specific lysis is shown as the mean of triplicate cultures at an E:T ratio of 5:1. (C) Transfection of UGT2B17 cDNA into donor B-LCL restores recognition by PL8 CTL. Donor B-LCL were transfected by electroporation with the full-length construct of UGT2B17 , selected for 3 d with puromycin (0.6 μg/ml), and assayed as targets for PL8 CTL. The lysis of UGT2B17 -transfected donor B-LCL (triangles), untransfected donor B-LCL (circles), and recipient B-LCL (squares) is shown at various E:T ratios.
    Human Stat3 Mrna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher pax6 against rodent mrna sequence
    UGT2B17 <t>mRNA</t> is transcribed in minor H antigen positive but not minor H antigen negative cells. (A) Northern blot analysis of total <t>RNA</t> from recipient and donor B-LCL, and from HLA-A29 + B-LCLs from 4 unrelated donors (designated 1–4). The blot was hybridized with a 32 P-labeled UGT2B17 probe for 16 h at 68°C. Detection of GAPDH mRNA was performed as a control. (B) Cytotoxicity assay for B-LCL by PL8 CTL. The B-LCL from unrelated donors ( 1 – 4 ) are the same lines used for analysis of gene expression in Fig. 3 A. Specific lysis is shown as the mean of triplicate cultures at an E:T ratio of 5:1. (C) Transfection of UGT2B17 cDNA into donor B-LCL restores recognition by PL8 CTL. Donor B-LCL were transfected by electroporation with the full-length construct of UGT2B17 , selected for 3 d with puromycin (0.6 μg/ml), and assayed as targets for PL8 CTL. The lysis of UGT2B17 -transfected donor B-LCL (triangles), untransfected donor B-LCL (circles), and recipient B-LCL (squares) is shown at various E:T ratios.
    Pax6 Against Rodent Mrna Sequence, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher genechip poly a rna control
    UGT2B17 <t>mRNA</t> is transcribed in minor H antigen positive but not minor H antigen negative cells. (A) Northern blot analysis of total <t>RNA</t> from recipient and donor B-LCL, and from HLA-A29 + B-LCLs from 4 unrelated donors (designated 1–4). The blot was hybridized with a 32 P-labeled UGT2B17 probe for 16 h at 68°C. Detection of GAPDH mRNA was performed as a control. (B) Cytotoxicity assay for B-LCL by PL8 CTL. The B-LCL from unrelated donors ( 1 – 4 ) are the same lines used for analysis of gene expression in Fig. 3 A. Specific lysis is shown as the mean of triplicate cultures at an E:T ratio of 5:1. (C) Transfection of UGT2B17 cDNA into donor B-LCL restores recognition by PL8 CTL. Donor B-LCL were transfected by electroporation with the full-length construct of UGT2B17 , selected for 3 d with puromycin (0.6 μg/ml), and assayed as targets for PL8 CTL. The lysis of UGT2B17 -transfected donor B-LCL (triangles), untransfected donor B-LCL (circles), and recipient B-LCL (squares) is shown at various E:T ratios.
    Genechip Poly A Rna Control, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 60 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher mrna isolation
    UGT2B17 <t>mRNA</t> is transcribed in minor H antigen positive but not minor H antigen negative cells. (A) Northern blot analysis of total <t>RNA</t> from recipient and donor B-LCL, and from HLA-A29 + B-LCLs from 4 unrelated donors (designated 1–4). The blot was hybridized with a 32 P-labeled UGT2B17 probe for 16 h at 68°C. Detection of GAPDH mRNA was performed as a control. (B) Cytotoxicity assay for B-LCL by PL8 CTL. The B-LCL from unrelated donors ( 1 – 4 ) are the same lines used for analysis of gene expression in Fig. 3 A. Specific lysis is shown as the mean of triplicate cultures at an E:T ratio of 5:1. (C) Transfection of UGT2B17 cDNA into donor B-LCL restores recognition by PL8 CTL. Donor B-LCL were transfected by electroporation with the full-length construct of UGT2B17 , selected for 3 d with puromycin (0.6 μg/ml), and assayed as targets for PL8 CTL. The lysis of UGT2B17 -transfected donor B-LCL (triangles), untransfected donor B-LCL (circles), and recipient B-LCL (squares) is shown at various E:T ratios.
    Mrna Isolation, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1943 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Thermo Fisher axin 1 messenger rna mrna
    (a) Dermal tissue messenger ribonucleic acid expressions of fibronectin, (b) transforming growth factor-beta 1, (c) Wnt II, and (d) <t>Axin-1.</t> Gene values were normalized to glyceraldehyde 3-phosphate dehydrogenase level; <t>mRNA:</t> Messenger ribonucleic acid; TGF: Transforming growth factor; † p value was
    Axin 1 Messenger Rna Mrna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher exogenous eukaryotic poly a rna
    (a) Dermal tissue messenger ribonucleic acid expressions of fibronectin, (b) transforming growth factor-beta 1, (c) Wnt II, and (d) <t>Axin-1.</t> Gene values were normalized to glyceraldehyde 3-phosphate dehydrogenase level; <t>mRNA:</t> Messenger ribonucleic acid; TGF: Transforming growth factor; † p value was
    Exogenous Eukaryotic Poly A Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Thermo Fisher chop messenger rna mrna
    (a) Dermal tissue messenger ribonucleic acid expressions of fibronectin, (b) transforming growth factor-beta 1, (c) Wnt II, and (d) <t>Axin-1.</t> Gene values were normalized to glyceraldehyde 3-phosphate dehydrogenase level; <t>mRNA:</t> Messenger ribonucleic acid; TGF: Transforming growth factor; † p value was
    Chop Messenger Rna Mrna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher mrna purification
    dPNUTS is a nuclear protein that colocalises with transcriptionally active RNAPII on salivary gland polytene chromosomes. A) Distribution of dPNUTS transcripts detected by <t>RNA</t> in situ hybridization; dPNUTS transcripts are maternally provided (top left) and are ubiquitously distributed in embryos at cellularisation (top right). At gastrulation, d PNUTS <t>mRNA</t> levels are enriched in the germband and in the fore- and hind-gut (fg and hg, respectively). Later, d PNUTS is highly expressed in the brain (br) and ventral nerve cord (vnc). Embryonic stage and approximate age, hours post fertilization (hpf), are indicated. B) 3 rd instar wing discs stained to reveal the distribution of ectopically expressed Myc-tagged dPNUTS (green in merge), Histone H3S10ph (red in merge, marking mitotic nuclei) and DNA. C) Images of whole mount salivary gland and magnified images of an individual nucleus (below), stained to show the localization of Myc-tagged dPNUTS (green in merge) and DNA (magenta in merge). D) Line scans of images in C) reveal that Myc-tagged dPNUTS is localised to interbands that stain weakly for DNA. Fluorescence intensity of anti-Myc antibody and TOPRO-3 staining was measured along a line through the indicated chromosomal region in the images shown. The profile plot below shows that the peaks of Myc-PNUTS and DNA of staining do not overlap. E) Polytene chromosomes from salivary gland squashes showing that dPNUTS localises to a number of discrete bands that are broadly distributed. F) Merging of the green signal representing dPNUTS with the red signal representing RNAPII Ser2-P (H5) identifies sites where these two proteins co-localize (example indicated with arrow). The relative signals of dPNUTS and RNAPII Ser2-P vary between sites, but the majority dPNUTS loci colocalize with RNAPII Ser2-P staining (star indicates example where only dPNUTS staining is visible).
    Mrna Purification, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 775 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Transfection of non-phagocytic cells, using mRNA:LPNs and mRNA:CS-PLGA at different ratios performed in epithelial A549 cells for 24 h and 48 h post-transfection using flow cytometer. mRNA complexed LPNs reveal a significant higher transgene expression over CS-PLGA NPs. Both NPs show higher transfection rates with higher mRNA:NP ratios and increasing transfection until 48 h post-transfection N = 4, mean ± SD (*p

    Journal: Journal of Nanobiotechnology

    Article Title: Kinetics of mRNA delivery and protein translation in dendritic cells using lipid-coated PLGA nanoparticles

    doi: 10.1186/s12951-018-0401-y

    Figure Lengend Snippet: Transfection of non-phagocytic cells, using mRNA:LPNs and mRNA:CS-PLGA at different ratios performed in epithelial A549 cells for 24 h and 48 h post-transfection using flow cytometer. mRNA complexed LPNs reveal a significant higher transgene expression over CS-PLGA NPs. Both NPs show higher transfection rates with higher mRNA:NP ratios and increasing transfection until 48 h post-transfection N = 4, mean ± SD (*p

    Article Snippet: The encapsulation efficiency (%EE) of bound mRNA:LPNs and mRNA:CS-PLGA NPs was evaluated indirectly by pelleting all samples down at 24,400g for 30 min and determining the concentration of unbound mRNA in the supernatant by measuring absorbance at 260/280 nm with a NanoDrop Spectrophotometer.

    Techniques: Transfection, Flow Cytometry, Cytometry, Expressing

    a1 Representative images depicted from live cell video after nine different time-points. The images are part of a video provided in Additional file 2 : Movie S1. DC2.4 cells were incubated with mRNA:FA-LPNs for a complete time duration of 4 h and with an interval of 3 min/image. Green dots on the images represent the fluorescence signal of labeled LPNs, while the cells signaling in red are the ones with successful mCherry expression. Scale bar = 100 µm. a2 Time-dependent change of the red fluorescence signal resulting from transfected cells and non-transfected, which are correlated with a3 green fluorescence signal of labeled nanoparticles. The tendency of each single cell being transfected is independent of the NPs uptake, as no significant difference between the uptake-behavior of transfected and non-transfected cells was observable. λ em = peak emission wavelength, MFI mean fluorescence intensity (mean ± SD, data from n = 32 fluorescent cells, n = 8 non-fluorescent cells, obtained from 4 independent videos)

    Journal: Journal of Nanobiotechnology

    Article Title: Kinetics of mRNA delivery and protein translation in dendritic cells using lipid-coated PLGA nanoparticles

    doi: 10.1186/s12951-018-0401-y

    Figure Lengend Snippet: a1 Representative images depicted from live cell video after nine different time-points. The images are part of a video provided in Additional file 2 : Movie S1. DC2.4 cells were incubated with mRNA:FA-LPNs for a complete time duration of 4 h and with an interval of 3 min/image. Green dots on the images represent the fluorescence signal of labeled LPNs, while the cells signaling in red are the ones with successful mCherry expression. Scale bar = 100 µm. a2 Time-dependent change of the red fluorescence signal resulting from transfected cells and non-transfected, which are correlated with a3 green fluorescence signal of labeled nanoparticles. The tendency of each single cell being transfected is independent of the NPs uptake, as no significant difference between the uptake-behavior of transfected and non-transfected cells was observable. λ em = peak emission wavelength, MFI mean fluorescence intensity (mean ± SD, data from n = 32 fluorescent cells, n = 8 non-fluorescent cells, obtained from 4 independent videos)

    Article Snippet: The encapsulation efficiency (%EE) of bound mRNA:LPNs and mRNA:CS-PLGA NPs was evaluated indirectly by pelleting all samples down at 24,400g for 30 min and determining the concentration of unbound mRNA in the supernatant by measuring absorbance at 260/280 nm with a NanoDrop Spectrophotometer.

    Techniques: Incubation, Fluorescence, Labeling, Expressing, Transfection

    Representative confocal images of DC2.4 cells transfected with both mRNA complexed NPs and by using jetPRIME ® as a positive control, naked mRNA as negative control. Transfection was analyzed with CLSM a 24 h and b 48 h post-transfection. Red fluorescence reveals cells successfully transfected with the nanoparticles while their morphology remains consistent with non-transfected cells (staining: green: cell membrane; blue: cell nucleus; scale bar: 50 μm)

    Journal: Journal of Nanobiotechnology

    Article Title: Kinetics of mRNA delivery and protein translation in dendritic cells using lipid-coated PLGA nanoparticles

    doi: 10.1186/s12951-018-0401-y

    Figure Lengend Snippet: Representative confocal images of DC2.4 cells transfected with both mRNA complexed NPs and by using jetPRIME ® as a positive control, naked mRNA as negative control. Transfection was analyzed with CLSM a 24 h and b 48 h post-transfection. Red fluorescence reveals cells successfully transfected with the nanoparticles while their morphology remains consistent with non-transfected cells (staining: green: cell membrane; blue: cell nucleus; scale bar: 50 μm)

    Article Snippet: The encapsulation efficiency (%EE) of bound mRNA:LPNs and mRNA:CS-PLGA NPs was evaluated indirectly by pelleting all samples down at 24,400g for 30 min and determining the concentration of unbound mRNA in the supernatant by measuring absorbance at 260/280 nm with a NanoDrop Spectrophotometer.

    Techniques: Transfection, Positive Control, Negative Control, Confocal Laser Scanning Microscopy, Fluorescence, Staining

    Quantification of cellular NP association for fluoresceinamine (FA) labeled blank ( a1 , a2 ) and mRNA complexed nanoparticles ( b1 , b2 ) tested in DC2.4 cells. NPs were incubated for 2 h and 4 h at different concentrations (20, 40 and 60 µg/mL corresponding to the weight ratios 1:10, 1:20 and 1:30 used for transfection). a1 , b1 Green NP fluorescent signal quantified by flow cytometry. Blank LPNs tend to show more uptake then blank CS-PLGA NPs, whereas for mRNA-loaded nanoparticles the opposite was observed. None of the samples showed a significant difference between 2 and 4 h incubation. a2 , b2 Representative graphs obtained for NP samples after 4 h incubation. N = 4, mean ± SD

    Journal: Journal of Nanobiotechnology

    Article Title: Kinetics of mRNA delivery and protein translation in dendritic cells using lipid-coated PLGA nanoparticles

    doi: 10.1186/s12951-018-0401-y

    Figure Lengend Snippet: Quantification of cellular NP association for fluoresceinamine (FA) labeled blank ( a1 , a2 ) and mRNA complexed nanoparticles ( b1 , b2 ) tested in DC2.4 cells. NPs were incubated for 2 h and 4 h at different concentrations (20, 40 and 60 µg/mL corresponding to the weight ratios 1:10, 1:20 and 1:30 used for transfection). a1 , b1 Green NP fluorescent signal quantified by flow cytometry. Blank LPNs tend to show more uptake then blank CS-PLGA NPs, whereas for mRNA-loaded nanoparticles the opposite was observed. None of the samples showed a significant difference between 2 and 4 h incubation. a2 , b2 Representative graphs obtained for NP samples after 4 h incubation. N = 4, mean ± SD

    Article Snippet: The encapsulation efficiency (%EE) of bound mRNA:LPNs and mRNA:CS-PLGA NPs was evaluated indirectly by pelleting all samples down at 24,400g for 30 min and determining the concentration of unbound mRNA in the supernatant by measuring absorbance at 260/280 nm with a NanoDrop Spectrophotometer.

    Techniques: Labeling, Incubation, Transfection, Flow Cytometry, Cytometry

    The kinetics of transfection for both mRNA-loaded NPs was quantified using flow cytometry, which indicated a significant difference between a1 mRNA:LPNs over a2 mRNA:CS-PLGA NPs, while mRNA:LPNs with a ratio of 1:20 and 1:30 elucidated a significant higher transfection rate over 1:10. a3 Control samples for transfection studies were JetPRIME ® as positive control and naked mRNA as negative control. (Note the enlarged y-axis, N = 4, mean ± SD.) a4 Representative graphs for evaluated time-points demonstrate a strong fluorescence shift for mRNA:LPNs with an increment in time and hence a higher transgene expression

    Journal: Journal of Nanobiotechnology

    Article Title: Kinetics of mRNA delivery and protein translation in dendritic cells using lipid-coated PLGA nanoparticles

    doi: 10.1186/s12951-018-0401-y

    Figure Lengend Snippet: The kinetics of transfection for both mRNA-loaded NPs was quantified using flow cytometry, which indicated a significant difference between a1 mRNA:LPNs over a2 mRNA:CS-PLGA NPs, while mRNA:LPNs with a ratio of 1:20 and 1:30 elucidated a significant higher transfection rate over 1:10. a3 Control samples for transfection studies were JetPRIME ® as positive control and naked mRNA as negative control. (Note the enlarged y-axis, N = 4, mean ± SD.) a4 Representative graphs for evaluated time-points demonstrate a strong fluorescence shift for mRNA:LPNs with an increment in time and hence a higher transgene expression

    Article Snippet: The encapsulation efficiency (%EE) of bound mRNA:LPNs and mRNA:CS-PLGA NPs was evaluated indirectly by pelleting all samples down at 24,400g for 30 min and determining the concentration of unbound mRNA in the supernatant by measuring absorbance at 260/280 nm with a NanoDrop Spectrophotometer.

    Techniques: Transfection, Flow Cytometry, Cytometry, Positive Control, Negative Control, Fluorescence, Expressing

    Schematic illustration of all nanoparticles used in this study. Both, LPNs and CS-PLGA NPs either complexed with mRNA or/and labeled with fluoresceinamine are used to quantify their uptake behavior and transfection efficiency in a dendritic cell line (DC2.4)

    Journal: Journal of Nanobiotechnology

    Article Title: Kinetics of mRNA delivery and protein translation in dendritic cells using lipid-coated PLGA nanoparticles

    doi: 10.1186/s12951-018-0401-y

    Figure Lengend Snippet: Schematic illustration of all nanoparticles used in this study. Both, LPNs and CS-PLGA NPs either complexed with mRNA or/and labeled with fluoresceinamine are used to quantify their uptake behavior and transfection efficiency in a dendritic cell line (DC2.4)

    Article Snippet: The encapsulation efficiency (%EE) of bound mRNA:LPNs and mRNA:CS-PLGA NPs was evaluated indirectly by pelleting all samples down at 24,400g for 30 min and determining the concentration of unbound mRNA in the supernatant by measuring absorbance at 260/280 nm with a NanoDrop Spectrophotometer.

    Techniques: Labeling, Transfection

    Gel retardation assay of mRNA complexed with different nanoparticles (NPs) at various ratios: a LPNs and b CS-PLGA NPs. Both images indicate mRNA binding to NPs and their appropriate release using heparin

    Journal: Journal of Nanobiotechnology

    Article Title: Kinetics of mRNA delivery and protein translation in dendritic cells using lipid-coated PLGA nanoparticles

    doi: 10.1186/s12951-018-0401-y

    Figure Lengend Snippet: Gel retardation assay of mRNA complexed with different nanoparticles (NPs) at various ratios: a LPNs and b CS-PLGA NPs. Both images indicate mRNA binding to NPs and their appropriate release using heparin

    Article Snippet: The encapsulation efficiency (%EE) of bound mRNA:LPNs and mRNA:CS-PLGA NPs was evaluated indirectly by pelleting all samples down at 24,400g for 30 min and determining the concentration of unbound mRNA in the supernatant by measuring absorbance at 260/280 nm with a NanoDrop Spectrophotometer.

    Techniques: Electrophoretic Mobility Shift Assay, Binding Assay

    Morphology of mRNA-loaded LPNs ( a1 , a2 ) and CS-PLGA NPs ( b1 , b2 ) visualized using SEM and TEM. a1 , b1 SEM images show a smooth, spherical morphology of the mRNA-loaded nanoparticles and a2 , b2 TEM images show the core–shell structure of the particles after staining with 0.5% (w/V) PTA

    Journal: Journal of Nanobiotechnology

    Article Title: Kinetics of mRNA delivery and protein translation in dendritic cells using lipid-coated PLGA nanoparticles

    doi: 10.1186/s12951-018-0401-y

    Figure Lengend Snippet: Morphology of mRNA-loaded LPNs ( a1 , a2 ) and CS-PLGA NPs ( b1 , b2 ) visualized using SEM and TEM. a1 , b1 SEM images show a smooth, spherical morphology of the mRNA-loaded nanoparticles and a2 , b2 TEM images show the core–shell structure of the particles after staining with 0.5% (w/V) PTA

    Article Snippet: The encapsulation efficiency (%EE) of bound mRNA:LPNs and mRNA:CS-PLGA NPs was evaluated indirectly by pelleting all samples down at 24,400g for 30 min and determining the concentration of unbound mRNA in the supernatant by measuring absorbance at 260/280 nm with a NanoDrop Spectrophotometer.

    Techniques: Transmission Electron Microscopy, Staining

    UGT2B17 mRNA is transcribed in minor H antigen positive but not minor H antigen negative cells. (A) Northern blot analysis of total RNA from recipient and donor B-LCL, and from HLA-A29 + B-LCLs from 4 unrelated donors (designated 1–4). The blot was hybridized with a 32 P-labeled UGT2B17 probe for 16 h at 68°C. Detection of GAPDH mRNA was performed as a control. (B) Cytotoxicity assay for B-LCL by PL8 CTL. The B-LCL from unrelated donors ( 1 – 4 ) are the same lines used for analysis of gene expression in Fig. 3 A. Specific lysis is shown as the mean of triplicate cultures at an E:T ratio of 5:1. (C) Transfection of UGT2B17 cDNA into donor B-LCL restores recognition by PL8 CTL. Donor B-LCL were transfected by electroporation with the full-length construct of UGT2B17 , selected for 3 d with puromycin (0.6 μg/ml), and assayed as targets for PL8 CTL. The lysis of UGT2B17 -transfected donor B-LCL (triangles), untransfected donor B-LCL (circles), and recipient B-LCL (squares) is shown at various E:T ratios.

    Journal: The Journal of Experimental Medicine

    Article Title: A Human Minor Histocompatibility Antigen Resulting from Differential Expression due to a Gene Deletion

    doi: 10.1084/jem.20030044

    Figure Lengend Snippet: UGT2B17 mRNA is transcribed in minor H antigen positive but not minor H antigen negative cells. (A) Northern blot analysis of total RNA from recipient and donor B-LCL, and from HLA-A29 + B-LCLs from 4 unrelated donors (designated 1–4). The blot was hybridized with a 32 P-labeled UGT2B17 probe for 16 h at 68°C. Detection of GAPDH mRNA was performed as a control. (B) Cytotoxicity assay for B-LCL by PL8 CTL. The B-LCL from unrelated donors ( 1 – 4 ) are the same lines used for analysis of gene expression in Fig. 3 A. Specific lysis is shown as the mean of triplicate cultures at an E:T ratio of 5:1. (C) Transfection of UGT2B17 cDNA into donor B-LCL restores recognition by PL8 CTL. Donor B-LCL were transfected by electroporation with the full-length construct of UGT2B17 , selected for 3 d with puromycin (0.6 μg/ml), and assayed as targets for PL8 CTL. The lysis of UGT2B17 -transfected donor B-LCL (triangles), untransfected donor B-LCL (circles), and recipient B-LCL (squares) is shown at various E:T ratios.

    Article Snippet: Total RNA was isolated from B-LCL and poly (A)+ mRNA was prepared by Straight A's mRNA Isolation (Novagen). mRNA was converted into cDNA using an oligo-dT primer that contains a NotI site at its 3′ end and Thermoscript reverse transcriptase (GIBCO BRL).

    Techniques: Northern Blot, Labeling, Cytotoxicity Assay, CTL Assay, Expressing, Lysis, Transfection, Electroporation, Construct

    (a) Dermal tissue messenger ribonucleic acid expressions of fibronectin, (b) transforming growth factor-beta 1, (c) Wnt II, and (d) Axin-1. Gene values were normalized to glyceraldehyde 3-phosphate dehydrogenase level; mRNA: Messenger ribonucleic acid; TGF: Transforming growth factor; † p value was

    Journal: Archives of Rheumatology

    Article Title: Paricalcitol Inhibits Wnt/β-Catenin Signaling Pathway and Ameliorates Dermal Fibrosis in Bleomycin Induced Scleroderma Model

    doi: 10.5606/ArchRheumatol.2018.6648

    Figure Lengend Snippet: (a) Dermal tissue messenger ribonucleic acid expressions of fibronectin, (b) transforming growth factor-beta 1, (c) Wnt II, and (d) Axin-1. Gene values were normalized to glyceraldehyde 3-phosphate dehydrogenase level; mRNA: Messenger ribonucleic acid; TGF: Transforming growth factor; † p value was

    Article Snippet: The samples were quantified for fibronectin, TGF- β1, Wnt-2 and axin-1 messenger RNA (mRNA) (Applied Biosystems, Foster City, CA, USA) using the comparative delta Ct, as described in the User’s Manual (Applied Biosystems, Foster City, CA, USA).

    Techniques:

    dPNUTS is a nuclear protein that colocalises with transcriptionally active RNAPII on salivary gland polytene chromosomes. A) Distribution of dPNUTS transcripts detected by RNA in situ hybridization; dPNUTS transcripts are maternally provided (top left) and are ubiquitously distributed in embryos at cellularisation (top right). At gastrulation, d PNUTS mRNA levels are enriched in the germband and in the fore- and hind-gut (fg and hg, respectively). Later, d PNUTS is highly expressed in the brain (br) and ventral nerve cord (vnc). Embryonic stage and approximate age, hours post fertilization (hpf), are indicated. B) 3 rd instar wing discs stained to reveal the distribution of ectopically expressed Myc-tagged dPNUTS (green in merge), Histone H3S10ph (red in merge, marking mitotic nuclei) and DNA. C) Images of whole mount salivary gland and magnified images of an individual nucleus (below), stained to show the localization of Myc-tagged dPNUTS (green in merge) and DNA (magenta in merge). D) Line scans of images in C) reveal that Myc-tagged dPNUTS is localised to interbands that stain weakly for DNA. Fluorescence intensity of anti-Myc antibody and TOPRO-3 staining was measured along a line through the indicated chromosomal region in the images shown. The profile plot below shows that the peaks of Myc-PNUTS and DNA of staining do not overlap. E) Polytene chromosomes from salivary gland squashes showing that dPNUTS localises to a number of discrete bands that are broadly distributed. F) Merging of the green signal representing dPNUTS with the red signal representing RNAPII Ser2-P (H5) identifies sites where these two proteins co-localize (example indicated with arrow). The relative signals of dPNUTS and RNAPII Ser2-P vary between sites, but the majority dPNUTS loci colocalize with RNAPII Ser2-P staining (star indicates example where only dPNUTS staining is visible).

    Journal: PLoS Genetics

    Article Title: PNUTS/PP1 Regulates RNAPII-Mediated Gene Expression and Is Necessary for Developmental Growth

    doi: 10.1371/journal.pgen.1003885

    Figure Lengend Snippet: dPNUTS is a nuclear protein that colocalises with transcriptionally active RNAPII on salivary gland polytene chromosomes. A) Distribution of dPNUTS transcripts detected by RNA in situ hybridization; dPNUTS transcripts are maternally provided (top left) and are ubiquitously distributed in embryos at cellularisation (top right). At gastrulation, d PNUTS mRNA levels are enriched in the germband and in the fore- and hind-gut (fg and hg, respectively). Later, d PNUTS is highly expressed in the brain (br) and ventral nerve cord (vnc). Embryonic stage and approximate age, hours post fertilization (hpf), are indicated. B) 3 rd instar wing discs stained to reveal the distribution of ectopically expressed Myc-tagged dPNUTS (green in merge), Histone H3S10ph (red in merge, marking mitotic nuclei) and DNA. C) Images of whole mount salivary gland and magnified images of an individual nucleus (below), stained to show the localization of Myc-tagged dPNUTS (green in merge) and DNA (magenta in merge). D) Line scans of images in C) reveal that Myc-tagged dPNUTS is localised to interbands that stain weakly for DNA. Fluorescence intensity of anti-Myc antibody and TOPRO-3 staining was measured along a line through the indicated chromosomal region in the images shown. The profile plot below shows that the peaks of Myc-PNUTS and DNA of staining do not overlap. E) Polytene chromosomes from salivary gland squashes showing that dPNUTS localises to a number of discrete bands that are broadly distributed. F) Merging of the green signal representing dPNUTS with the red signal representing RNAPII Ser2-P (H5) identifies sites where these two proteins co-localize (example indicated with arrow). The relative signals of dPNUTS and RNAPII Ser2-P vary between sites, but the majority dPNUTS loci colocalize with RNAPII Ser2-P staining (star indicates example where only dPNUTS staining is visible).

    Article Snippet: Total RNA quality and quantity was verified on a NanoDrop1000 spectrophotometer (Thermofisher) and Bioanalyzer 2100. mRNA was polyA selected using Dynabeads mRNA Purification Kit for mRNA Purification from Total RNA Preps (Invitrogen).

    Techniques: RNA In Situ Hybridization, Staining, Fluorescence