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m6atp  (Jena Bioscience)


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    Structured Review

    Jena Bioscience m6atp
    The NlucP reporter mRNAs with 5’ and 3’ UTRs of SLU7 , RPL32, and HSP70 were transcribed in vitro in the presence of 50% or 0% m⁶ATP. mRNAs were then enzymatically capped and polyadenylated (A-B) . For the pull-down assay (C) , after transcription, the mRNAs were first polyadenylated, then biotinylated at the 3’ end, and enzymatically capped. (A) mRNAs were translated in CFTS prepared from untreated HEK293T cells in the presence of 0.1 mM cap-analogs m7GpppG or ApppG. NlucP activity in the presence of m7GpppG was normalized to that in the presence of ApppG. (B) mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. NlucP activity in Torin1-treated CFTS was normalized to that in DMSO-treated CFTS. Values are the means of at least three independent replicates. Error bars show standard deviations. Two-tailed Student’s t-test was used to estimate the statistical significance between methylated and unmethylated mRNAs. ***p<0.001. (C) The biotinylated mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. RNA-protein complexes were isolated using streptavidin-sepharose and analysed by Western blot using antibodies against eIF4A, eIF4E, and eIF4G. Uncapped mRNA with RPL32 UTRs (K-) was used as a control. CFTS: cell-free translation system; RLU: relative luciferase units.
    M6atp, supplied by Jena Bioscience, used in various techniques. Bioz Stars score: 93/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    m6atp - by Bioz Stars, 2026-03
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    1) Product Images from "Transcript-wide m 6 A methylation defines the efficiency of the cap-independent translation initiation"

    Article Title: Transcript-wide m 6 A methylation defines the efficiency of the cap-independent translation initiation

    Journal: bioRxiv

    doi: 10.1101/2025.11.25.690160

    The NlucP reporter mRNAs with 5’ and 3’ UTRs of SLU7 , RPL32, and HSP70 were transcribed in vitro in the presence of 50% or 0% m⁶ATP. mRNAs were then enzymatically capped and polyadenylated (A-B) . For the pull-down assay (C) , after transcription, the mRNAs were first polyadenylated, then biotinylated at the 3’ end, and enzymatically capped. (A) mRNAs were translated in CFTS prepared from untreated HEK293T cells in the presence of 0.1 mM cap-analogs m7GpppG or ApppG. NlucP activity in the presence of m7GpppG was normalized to that in the presence of ApppG. (B) mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. NlucP activity in Torin1-treated CFTS was normalized to that in DMSO-treated CFTS. Values are the means of at least three independent replicates. Error bars show standard deviations. Two-tailed Student’s t-test was used to estimate the statistical significance between methylated and unmethylated mRNAs. ***p<0.001. (C) The biotinylated mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. RNA-protein complexes were isolated using streptavidin-sepharose and analysed by Western blot using antibodies against eIF4A, eIF4E, and eIF4G. Uncapped mRNA with RPL32 UTRs (K-) was used as a control. CFTS: cell-free translation system; RLU: relative luciferase units.
    Figure Legend Snippet: The NlucP reporter mRNAs with 5’ and 3’ UTRs of SLU7 , RPL32, and HSP70 were transcribed in vitro in the presence of 50% or 0% m⁶ATP. mRNAs were then enzymatically capped and polyadenylated (A-B) . For the pull-down assay (C) , after transcription, the mRNAs were first polyadenylated, then biotinylated at the 3’ end, and enzymatically capped. (A) mRNAs were translated in CFTS prepared from untreated HEK293T cells in the presence of 0.1 mM cap-analogs m7GpppG or ApppG. NlucP activity in the presence of m7GpppG was normalized to that in the presence of ApppG. (B) mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. NlucP activity in Torin1-treated CFTS was normalized to that in DMSO-treated CFTS. Values are the means of at least three independent replicates. Error bars show standard deviations. Two-tailed Student’s t-test was used to estimate the statistical significance between methylated and unmethylated mRNAs. ***p<0.001. (C) The biotinylated mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. RNA-protein complexes were isolated using streptavidin-sepharose and analysed by Western blot using antibodies against eIF4A, eIF4E, and eIF4G. Uncapped mRNA with RPL32 UTRs (K-) was used as a control. CFTS: cell-free translation system; RLU: relative luciferase units.

    Techniques Used: In Vitro, Pull Down Assay, Activity Assay, Two Tailed Test, Methylation, Isolation, Western Blot, Control, Luciferase

    The barplots show the relative luciferase activity for the RPL32 reporter mRNAs transcribed in vitro with the indicated m⁶ATP percentage (X-axis). (A) mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. NlucP activity in CFTS obtained from Torin1-treated cells was normalized to that in the CFTS from DMSO-treated cells. (B) mRNA transfected into WT HEK293T cells, pre-treated for 2h by 250nM Torin1 or DMSO. The mRNAs were transfected together with furimazine, and luciferase activity was measured in living cells at 40 min for the CFTS assay and at 80 min for the ex vivo assay. NlucP activity in Torin1-treated cells was normalized to that in DMSO-treated cells. Values are the means of at least three independent replicates. The error bars show standard deviations. The results were analyzed using the two-tailed Student’s t-test, with the significance levels indicated as follows: *p < 0.05; **p < 0.01; ***p < 0.001.
    Figure Legend Snippet: The barplots show the relative luciferase activity for the RPL32 reporter mRNAs transcribed in vitro with the indicated m⁶ATP percentage (X-axis). (A) mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. NlucP activity in CFTS obtained from Torin1-treated cells was normalized to that in the CFTS from DMSO-treated cells. (B) mRNA transfected into WT HEK293T cells, pre-treated for 2h by 250nM Torin1 or DMSO. The mRNAs were transfected together with furimazine, and luciferase activity was measured in living cells at 40 min for the CFTS assay and at 80 min for the ex vivo assay. NlucP activity in Torin1-treated cells was normalized to that in DMSO-treated cells. Values are the means of at least three independent replicates. The error bars show standard deviations. The results were analyzed using the two-tailed Student’s t-test, with the significance levels indicated as follows: *p < 0.05; **p < 0.01; ***p < 0.001.

    Techniques Used: Luciferase, Activity Assay, In Vitro, Transfection, Ex Vivo, Two Tailed Test



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    Image Search Results


    The NlucP reporter mRNAs with 5’ and 3’ UTRs of SLU7 , RPL32, and HSP70 were transcribed in vitro in the presence of 50% or 0% m⁶ATP. mRNAs were then enzymatically capped and polyadenylated (A-B) . For the pull-down assay (C) , after transcription, the mRNAs were first polyadenylated, then biotinylated at the 3’ end, and enzymatically capped. (A) mRNAs were translated in CFTS prepared from untreated HEK293T cells in the presence of 0.1 mM cap-analogs m7GpppG or ApppG. NlucP activity in the presence of m7GpppG was normalized to that in the presence of ApppG. (B) mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. NlucP activity in Torin1-treated CFTS was normalized to that in DMSO-treated CFTS. Values are the means of at least three independent replicates. Error bars show standard deviations. Two-tailed Student’s t-test was used to estimate the statistical significance between methylated and unmethylated mRNAs. ***p<0.001. (C) The biotinylated mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. RNA-protein complexes were isolated using streptavidin-sepharose and analysed by Western blot using antibodies against eIF4A, eIF4E, and eIF4G. Uncapped mRNA with RPL32 UTRs (K-) was used as a control. CFTS: cell-free translation system; RLU: relative luciferase units.

    Journal: bioRxiv

    Article Title: Transcript-wide m 6 A methylation defines the efficiency of the cap-independent translation initiation

    doi: 10.1101/2025.11.25.690160

    Figure Lengend Snippet: The NlucP reporter mRNAs with 5’ and 3’ UTRs of SLU7 , RPL32, and HSP70 were transcribed in vitro in the presence of 50% or 0% m⁶ATP. mRNAs were then enzymatically capped and polyadenylated (A-B) . For the pull-down assay (C) , after transcription, the mRNAs were first polyadenylated, then biotinylated at the 3’ end, and enzymatically capped. (A) mRNAs were translated in CFTS prepared from untreated HEK293T cells in the presence of 0.1 mM cap-analogs m7GpppG or ApppG. NlucP activity in the presence of m7GpppG was normalized to that in the presence of ApppG. (B) mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. NlucP activity in Torin1-treated CFTS was normalized to that in DMSO-treated CFTS. Values are the means of at least three independent replicates. Error bars show standard deviations. Two-tailed Student’s t-test was used to estimate the statistical significance between methylated and unmethylated mRNAs. ***p<0.001. (C) The biotinylated mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. RNA-protein complexes were isolated using streptavidin-sepharose and analysed by Western blot using antibodies against eIF4A, eIF4E, and eIF4G. Uncapped mRNA with RPL32 UTRs (K-) was used as a control. CFTS: cell-free translation system; RLU: relative luciferase units.

    Article Snippet: In vitro transcription was performed using the T7-Scribe Standard RNA IVT Kit (CellScript, USA) according to the manufacturer’s instructions. m6A-Methylated mRNAs were generated by incorporating m6ATP (Jena Bioscience, Germany, NU-1101L) along with ATP at defined ratios.

    Techniques: In Vitro, Pull Down Assay, Activity Assay, Two Tailed Test, Methylation, Isolation, Western Blot, Control, Luciferase

    The barplots show the relative luciferase activity for the RPL32 reporter mRNAs transcribed in vitro with the indicated m⁶ATP percentage (X-axis). (A) mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. NlucP activity in CFTS obtained from Torin1-treated cells was normalized to that in the CFTS from DMSO-treated cells. (B) mRNA transfected into WT HEK293T cells, pre-treated for 2h by 250nM Torin1 or DMSO. The mRNAs were transfected together with furimazine, and luciferase activity was measured in living cells at 40 min for the CFTS assay and at 80 min for the ex vivo assay. NlucP activity in Torin1-treated cells was normalized to that in DMSO-treated cells. Values are the means of at least three independent replicates. The error bars show standard deviations. The results were analyzed using the two-tailed Student’s t-test, with the significance levels indicated as follows: *p < 0.05; **p < 0.01; ***p < 0.001.

    Journal: bioRxiv

    Article Title: Transcript-wide m 6 A methylation defines the efficiency of the cap-independent translation initiation

    doi: 10.1101/2025.11.25.690160

    Figure Lengend Snippet: The barplots show the relative luciferase activity for the RPL32 reporter mRNAs transcribed in vitro with the indicated m⁶ATP percentage (X-axis). (A) mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. NlucP activity in CFTS obtained from Torin1-treated cells was normalized to that in the CFTS from DMSO-treated cells. (B) mRNA transfected into WT HEK293T cells, pre-treated for 2h by 250nM Torin1 or DMSO. The mRNAs were transfected together with furimazine, and luciferase activity was measured in living cells at 40 min for the CFTS assay and at 80 min for the ex vivo assay. NlucP activity in Torin1-treated cells was normalized to that in DMSO-treated cells. Values are the means of at least three independent replicates. The error bars show standard deviations. The results were analyzed using the two-tailed Student’s t-test, with the significance levels indicated as follows: *p < 0.05; **p < 0.01; ***p < 0.001.

    Article Snippet: In vitro transcription was performed using the T7-Scribe Standard RNA IVT Kit (CellScript, USA) according to the manufacturer’s instructions. m6A-Methylated mRNAs were generated by incorporating m6ATP (Jena Bioscience, Germany, NU-1101L) along with ATP at defined ratios.

    Techniques: Luciferase, Activity Assay, In Vitro, Transfection, Ex Vivo, Two Tailed Test