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polya mrna selection  (New England Biolabs)


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    New England Biolabs polya mrna selection
    (A) Volcano plot analysis of host transcriptome-wide genes differentially expressed in primary B cells from three independent donors infected with WT or TES2m LMP1 for 21 days. Positive fold changes indicate higher transcripts in cells with TES2m LMP1. (B) PFKFB4 glucose metabolism roles. Glucose is metabolized by glycolysis or by the PPP. Glycolysis flux is regulated via Fructose-2,6-BP, an allosteric regulator of PFK1. PFKFB3 phosphorylates Fructose-6-P into Fructose-2,6-BP while PFKFB4 phosphatase activity converts Fructose-2,6-BP to Fructose-6-P to shunt glycolytic flux to PPP. Created in BioRender. Burton, E. (2026) https://BioRender.com/i0gawc8 . (C) KEGG pathway analysis of transcripts significantly upregulated in cells infected by EBV with WT LMP1 at day 21 post infection. (D) KEGG pathway analysis of transcripts significantly upregulated in cells infected by TES2m EBV at day 21 post infection. (E) PFKFB4 protein levels in WT versus TES2m LMP1 EBV infected cells. Cells from two independent donors were infected with EBV with WT LMP1 or TES2m LMP1 for 22 days. Densitometry values of PFKFB4 vs β-Actin load controls are shown. (F) Effects of LCL LMP1 KO on PFKFB4 <t>mRNA.</t> Median PFKFB4 reads from RNAseq analysis of Cas9+ GM12878 cells with control vs LMP1 targeting sgRNA expression for 48 hours. Shown are data from n=3 RNAseq datasets. (G) PFKFB4 protein levels in LMP1 KO LCLs. Cas9+ GM12878 were mock induced or induced to express LMP1 sgRNA for 48 hours. WCL was extracted and immunoblot was carried out using the indicated antibodies. (H) Effects of TES1 vs TES2 signaling inhibition on LCL PFKFB4 expression. Median PFKFB4 RNAseq reads from RNAseq analysis of Cas9+ GM12878 expressing LMP1 sgRNA together with dox-induced LMP1 TES1m vs WT LMP1 cDNA expression. Shown are data from n=3 RNAseq datasets. (I) Relative PFKFB4 protein abundances from proteomic analysis of primary human B-cells at the indicated days post-infection by the EBV B95.8 strain. Shown are mean ± SD values from n=4 replicates. (J) Effects of exogenous PFKFB4 expression on proliferation of primary B cells infected by EBV with WT versus TES2m LMP1. Peripheral blood B cells from three independent donors were infected with EBV with WT vs TES2m LMP1. After 14 days, cells were transduced with lentivirus control or with lentivirus driving stable PFKFB4 expression. Five days post-selection, cells were re-seeded. Twelve days later, live cell counts were defined. P-values were determined by one-sided Fisher’s exact test. * p<0.05, **p<0.005, ***p<0.0005. blots are representative of n=3 independent replicates.
    Polya Mrna Selection, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 6312 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/polya+mrna+selection/NEBNext+Poly(A)+mRNA+Magnetic+Isolation+Module/bio_rxiv__64898__2026__03__09__710628-277-12-16
    Average 99 stars, based on 6312 article reviews
    polya mrna selection - by Bioz Stars, 2026-09
    99/100 stars

    Images

    1) Product Images from "Epstein-Barr Virus Latent Membrane Protein 1 Suppresses Ferroptosis via Pentose Phosphate Pathway and Glutathione Metabolism"

    Article Title: Epstein-Barr Virus Latent Membrane Protein 1 Suppresses Ferroptosis via Pentose Phosphate Pathway and Glutathione Metabolism

    Journal: bioRxiv

    doi: 10.64898/2026.03.09.710628

    (A) Volcano plot analysis of host transcriptome-wide genes differentially expressed in primary B cells from three independent donors infected with WT or TES2m LMP1 for 21 days. Positive fold changes indicate higher transcripts in cells with TES2m LMP1. (B) PFKFB4 glucose metabolism roles. Glucose is metabolized by glycolysis or by the PPP. Glycolysis flux is regulated via Fructose-2,6-BP, an allosteric regulator of PFK1. PFKFB3 phosphorylates Fructose-6-P into Fructose-2,6-BP while PFKFB4 phosphatase activity converts Fructose-2,6-BP to Fructose-6-P to shunt glycolytic flux to PPP. Created in BioRender. Burton, E. (2026) https://BioRender.com/i0gawc8 . (C) KEGG pathway analysis of transcripts significantly upregulated in cells infected by EBV with WT LMP1 at day 21 post infection. (D) KEGG pathway analysis of transcripts significantly upregulated in cells infected by TES2m EBV at day 21 post infection. (E) PFKFB4 protein levels in WT versus TES2m LMP1 EBV infected cells. Cells from two independent donors were infected with EBV with WT LMP1 or TES2m LMP1 for 22 days. Densitometry values of PFKFB4 vs β-Actin load controls are shown. (F) Effects of LCL LMP1 KO on PFKFB4 mRNA. Median PFKFB4 reads from RNAseq analysis of Cas9+ GM12878 cells with control vs LMP1 targeting sgRNA expression for 48 hours. Shown are data from n=3 RNAseq datasets. (G) PFKFB4 protein levels in LMP1 KO LCLs. Cas9+ GM12878 were mock induced or induced to express LMP1 sgRNA for 48 hours. WCL was extracted and immunoblot was carried out using the indicated antibodies. (H) Effects of TES1 vs TES2 signaling inhibition on LCL PFKFB4 expression. Median PFKFB4 RNAseq reads from RNAseq analysis of Cas9+ GM12878 expressing LMP1 sgRNA together with dox-induced LMP1 TES1m vs WT LMP1 cDNA expression. Shown are data from n=3 RNAseq datasets. (I) Relative PFKFB4 protein abundances from proteomic analysis of primary human B-cells at the indicated days post-infection by the EBV B95.8 strain. Shown are mean ± SD values from n=4 replicates. (J) Effects of exogenous PFKFB4 expression on proliferation of primary B cells infected by EBV with WT versus TES2m LMP1. Peripheral blood B cells from three independent donors were infected with EBV with WT vs TES2m LMP1. After 14 days, cells were transduced with lentivirus control or with lentivirus driving stable PFKFB4 expression. Five days post-selection, cells were re-seeded. Twelve days later, live cell counts were defined. P-values were determined by one-sided Fisher’s exact test. * p<0.05, **p<0.005, ***p<0.0005. blots are representative of n=3 independent replicates.
    Figure Legend Snippet: (A) Volcano plot analysis of host transcriptome-wide genes differentially expressed in primary B cells from three independent donors infected with WT or TES2m LMP1 for 21 days. Positive fold changes indicate higher transcripts in cells with TES2m LMP1. (B) PFKFB4 glucose metabolism roles. Glucose is metabolized by glycolysis or by the PPP. Glycolysis flux is regulated via Fructose-2,6-BP, an allosteric regulator of PFK1. PFKFB3 phosphorylates Fructose-6-P into Fructose-2,6-BP while PFKFB4 phosphatase activity converts Fructose-2,6-BP to Fructose-6-P to shunt glycolytic flux to PPP. Created in BioRender. Burton, E. (2026) https://BioRender.com/i0gawc8 . (C) KEGG pathway analysis of transcripts significantly upregulated in cells infected by EBV with WT LMP1 at day 21 post infection. (D) KEGG pathway analysis of transcripts significantly upregulated in cells infected by TES2m EBV at day 21 post infection. (E) PFKFB4 protein levels in WT versus TES2m LMP1 EBV infected cells. Cells from two independent donors were infected with EBV with WT LMP1 or TES2m LMP1 for 22 days. Densitometry values of PFKFB4 vs β-Actin load controls are shown. (F) Effects of LCL LMP1 KO on PFKFB4 mRNA. Median PFKFB4 reads from RNAseq analysis of Cas9+ GM12878 cells with control vs LMP1 targeting sgRNA expression for 48 hours. Shown are data from n=3 RNAseq datasets. (G) PFKFB4 protein levels in LMP1 KO LCLs. Cas9+ GM12878 were mock induced or induced to express LMP1 sgRNA for 48 hours. WCL was extracted and immunoblot was carried out using the indicated antibodies. (H) Effects of TES1 vs TES2 signaling inhibition on LCL PFKFB4 expression. Median PFKFB4 RNAseq reads from RNAseq analysis of Cas9+ GM12878 expressing LMP1 sgRNA together with dox-induced LMP1 TES1m vs WT LMP1 cDNA expression. Shown are data from n=3 RNAseq datasets. (I) Relative PFKFB4 protein abundances from proteomic analysis of primary human B-cells at the indicated days post-infection by the EBV B95.8 strain. Shown are mean ± SD values from n=4 replicates. (J) Effects of exogenous PFKFB4 expression on proliferation of primary B cells infected by EBV with WT versus TES2m LMP1. Peripheral blood B cells from three independent donors were infected with EBV with WT vs TES2m LMP1. After 14 days, cells were transduced with lentivirus control or with lentivirus driving stable PFKFB4 expression. Five days post-selection, cells were re-seeded. Twelve days later, live cell counts were defined. P-values were determined by one-sided Fisher’s exact test. * p<0.05, **p<0.005, ***p<0.0005. blots are representative of n=3 independent replicates.

    Techniques Used: Infection, Activity Assay, RNA sequencing, Control, Expressing, Western Blot, Inhibition, Transduction, Selection

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    Article Title: Characterization of Target Gene Regulation by the Two Epstein-Barr Virus Oncogene LMP1 Domains Essential for B-cell Transformation
    Article Snippet: Total RNA was isolated using RNeasy mini kit (Qiagen #74106) with in-column genomic DNA digestion step according to the manufacturer’s protocol. .. To construct indexed libraries, 1 μg of total RNA was used for polyA mRNA selection using NEBNext Poly(A) mRNA Magnetic Isolation Module (Cat#E7490S), and library preparation with NEBNext Ultra RNA Library Prep with Sample Purification Beads (Cat#E7765S). ..

    Article Title: Characterization of target gene regulation by the two Epstein-Barr virus oncogene LMP1 domains essential for B-cell transformation
    Article Snippet: Total RNA was isolated using RNeasy mini kit (Qiagen #74106) with in-column genomic DNA digestion step (RNase-free DNase set, Qiagen #79254) according to the manufacturer’s protocol. .. To construct indexed libraries, 1 μg of total RNA was used for polyA mRNA selection using NEBNext Poly(A) mRNA Magnetic Isolation Module (Cat#E7490S), and library preparation with NEBNext Ultra RNA Library Prep with Sample Purification Beads (Cat#E7765S). ..

    Article Title: Epstein-Barr virus-transformed B-cells from a Hypoxia model of the germinal center requires external unsaturated fatty acids.
    Article Snippet: .. To construct indexed libraries, 1 μg of total RNA was used for polyA mRNA-selection, using the NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs), followed by library construction via the NEBNext Ultra RNA Library Prep Kit (New England Biolabs). ..

    Article Title: Epstein-Barr Virus Latent Membrane Protein 1 Suppresses Ferroptosis via Pentose Phosphate Pathway and Glutathione Metabolism
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    Article Title: Epstein-Barr virus-transformed B-cells from a hypoxia model of the germinal center requires external unsaturated fatty acids
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    Selection:

    Article Title: Characterization of Target Gene Regulation by the Two Epstein-Barr Virus Oncogene LMP1 Domains Essential for B-cell Transformation
    Article Snippet: Total RNA was isolated using RNeasy mini kit (Qiagen #74106) with in-column genomic DNA digestion step according to the manufacturer’s protocol. .. To construct indexed libraries, 1 μg of total RNA was used for polyA mRNA selection using NEBNext Poly(A) mRNA Magnetic Isolation Module (Cat#E7490S), and library preparation with NEBNext Ultra RNA Library Prep with Sample Purification Beads (Cat#E7765S). ..

    Article Title: Characterization of target gene regulation by the two Epstein-Barr virus oncogene LMP1 domains essential for B-cell transformation
    Article Snippet: Total RNA was isolated using RNeasy mini kit (Qiagen #74106) with in-column genomic DNA digestion step (RNase-free DNase set, Qiagen #79254) according to the manufacturer’s protocol. .. To construct indexed libraries, 1 μg of total RNA was used for polyA mRNA selection using NEBNext Poly(A) mRNA Magnetic Isolation Module (Cat#E7490S), and library preparation with NEBNext Ultra RNA Library Prep with Sample Purification Beads (Cat#E7765S). ..

    Article Title: Epstein-Barr Virus Latent Membrane Protein 1 Suppresses Ferroptosis via Pentose Phosphate Pathway and Glutathione Metabolism
    Article Snippet: Total RNA was isolated using RNeasy mini kit (Qiagen #74106) with in-column genomic DNA digestion step (RNase-free DNase set, Qiagen #79254) according to the manufacturer’s protocol. .. To construct indexed libraries, 1 μg of total RNA was used for polyA mRNA selection using NEBNext Poly(A) mRNA Magnetic Isolation Module (Cat#E7490S), and library preparation with NEBNext Ultra RNA Library Prep with Sample Purification Beads (Cat#E7765S). ..

    Isolation:

    Article Title: Characterization of Target Gene Regulation by the Two Epstein-Barr Virus Oncogene LMP1 Domains Essential for B-cell Transformation
    Article Snippet: Total RNA was isolated using RNeasy mini kit (Qiagen #74106) with in-column genomic DNA digestion step according to the manufacturer’s protocol. .. To construct indexed libraries, 1 μg of total RNA was used for polyA mRNA selection using NEBNext Poly(A) mRNA Magnetic Isolation Module (Cat#E7490S), and library preparation with NEBNext Ultra RNA Library Prep with Sample Purification Beads (Cat#E7765S). ..

    Article Title: Characterization of target gene regulation by the two Epstein-Barr virus oncogene LMP1 domains essential for B-cell transformation
    Article Snippet: Total RNA was isolated using RNeasy mini kit (Qiagen #74106) with in-column genomic DNA digestion step (RNase-free DNase set, Qiagen #79254) according to the manufacturer’s protocol. .. To construct indexed libraries, 1 μg of total RNA was used for polyA mRNA selection using NEBNext Poly(A) mRNA Magnetic Isolation Module (Cat#E7490S), and library preparation with NEBNext Ultra RNA Library Prep with Sample Purification Beads (Cat#E7765S). ..

    Article Title: Epstein-Barr virus-transformed B-cells from a Hypoxia model of the germinal center requires external unsaturated fatty acids.
    Article Snippet: .. To construct indexed libraries, 1 μg of total RNA was used for polyA mRNA-selection, using the NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs), followed by library construction via the NEBNext Ultra RNA Library Prep Kit (New England Biolabs). ..

    Article Title: Epstein-Barr Virus Latent Membrane Protein 1 Suppresses Ferroptosis via Pentose Phosphate Pathway and Glutathione Metabolism
    Article Snippet: Total RNA was isolated using RNeasy mini kit (Qiagen #74106) with in-column genomic DNA digestion step (RNase-free DNase set, Qiagen #79254) according to the manufacturer’s protocol. .. To construct indexed libraries, 1 μg of total RNA was used for polyA mRNA selection using NEBNext Poly(A) mRNA Magnetic Isolation Module (Cat#E7490S), and library preparation with NEBNext Ultra RNA Library Prep with Sample Purification Beads (Cat#E7765S). ..

    Article Title: Methionine metabolism controls the B cell EBV epigenome and viral latency.
    Article Snippet: To construct indexed libraries, 1 mg of total RNA was used for polyA mRNA-selection, using the NEBNext Poly(A) mRNAMagnetic Isolation Module (New England Biolabs), followed by library construction via the NEBNext Ultra RNA Library Prep Kit (New England Biolabs). .. To construct indexed libraries, 1 mg of total RNA was used for polyA mRNA-selection, using the NEBNext Poly(A) mRNAMagnetic Isolation Module (New England Biolabs), followed by library construction via the NEBNext Ultra RNA Library Prep Kit (New England Biolabs). .. Libraries were multi-indexed, pooled and sequenced on an Illumina NextSeq 500 sequencer using single-end 75 bp reads (Illunima) at the Dana Farber Molecular Biology core.

    Article Title: Multi-omic Analysis of Human B-cell Activation Reveals a Key Lysosomal BCAT1 Role in mTOR Hyperactivation by B-cell receptor and TLR9
    Article Snippet: .. To construct indexed libraries, 1 μg of total RNA was used for polyA mRNA-selection, using the NEBNext Page 18/44 Poly(A) mRNA Magnetic Isolation Module (New England Biolabs), followed by library construction via the NEBNext Ultra RNA Library Prep Kit (New England Biolabs). ..

    Article Title: Epstein-Barr virus-transformed B-cells from a hypoxia model of the germinal center requires external unsaturated fatty acids
    Article Snippet: .. To construct indexed libraries, 1 μg of total RNA was used for polyA mRNA-selection, using the NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs), followed by library construction via the NEBNext Ultra RNA Library Prep Kit (New England Biolabs). ..

    Article Title: Epstein-Barr virus BNRF1 destabilizes SMC5/6 cohesin complexes to evade its restriction of replication compartments
    Article Snippet: .. To construct indexed libraries, 1 μg of total RNA was used for polyA mRNA-selection using NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB), followed by library construction using NEBNext Ultra RNA Library Prep Kit for Illumina (NEB). ..

    Sample Purification:

    Article Title: Characterization of Target Gene Regulation by the Two Epstein-Barr Virus Oncogene LMP1 Domains Essential for B-cell Transformation
    Article Snippet: Total RNA was isolated using RNeasy mini kit (Qiagen #74106) with in-column genomic DNA digestion step according to the manufacturer’s protocol. .. To construct indexed libraries, 1 μg of total RNA was used for polyA mRNA selection using NEBNext Poly(A) mRNA Magnetic Isolation Module (Cat#E7490S), and library preparation with NEBNext Ultra RNA Library Prep with Sample Purification Beads (Cat#E7765S). ..

    Article Title: Characterization of target gene regulation by the two Epstein-Barr virus oncogene LMP1 domains essential for B-cell transformation
    Article Snippet: Total RNA was isolated using RNeasy mini kit (Qiagen #74106) with in-column genomic DNA digestion step (RNase-free DNase set, Qiagen #79254) according to the manufacturer’s protocol. .. To construct indexed libraries, 1 μg of total RNA was used for polyA mRNA selection using NEBNext Poly(A) mRNA Magnetic Isolation Module (Cat#E7490S), and library preparation with NEBNext Ultra RNA Library Prep with Sample Purification Beads (Cat#E7765S). ..

    Article Title: Epstein-Barr Virus Latent Membrane Protein 1 Suppresses Ferroptosis via Pentose Phosphate Pathway and Glutathione Metabolism
    Article Snippet: Total RNA was isolated using RNeasy mini kit (Qiagen #74106) with in-column genomic DNA digestion step (RNase-free DNase set, Qiagen #79254) according to the manufacturer’s protocol. .. To construct indexed libraries, 1 μg of total RNA was used for polyA mRNA selection using NEBNext Poly(A) mRNA Magnetic Isolation Module (Cat#E7490S), and library preparation with NEBNext Ultra RNA Library Prep with Sample Purification Beads (Cat#E7765S). ..



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    (A) Volcano plot analysis of host transcriptome-wide genes differentially expressed in primary B cells from three independent donors infected with WT or TES2m LMP1 for 21 days. Positive fold changes indicate higher transcripts in cells with TES2m LMP1. (B) PFKFB4 glucose metabolism roles. Glucose is metabolized by glycolysis or by the PPP. Glycolysis flux is regulated via Fructose-2,6-BP, an allosteric regulator of PFK1. PFKFB3 phosphorylates Fructose-6-P into Fructose-2,6-BP while PFKFB4 phosphatase activity converts Fructose-2,6-BP to Fructose-6-P to shunt glycolytic flux to PPP. Created in BioRender. Burton, E. (2026) https://BioRender.com/i0gawc8 . (C) KEGG pathway analysis of transcripts significantly upregulated in cells infected by EBV with WT LMP1 at day 21 post infection. (D) KEGG pathway analysis of transcripts significantly upregulated in cells infected by TES2m EBV at day 21 post infection. (E) PFKFB4 protein levels in WT versus TES2m LMP1 EBV infected cells. Cells from two independent donors were infected with EBV with WT LMP1 or TES2m LMP1 for 22 days. Densitometry values of PFKFB4 vs β-Actin load controls are shown. (F) Effects of LCL LMP1 KO on PFKFB4 <t>mRNA.</t> Median PFKFB4 reads from RNAseq analysis of Cas9+ GM12878 cells with control vs LMP1 targeting sgRNA expression for 48 hours. Shown are data from n=3 RNAseq datasets. (G) PFKFB4 protein levels in LMP1 KO LCLs. Cas9+ GM12878 were mock induced or induced to express LMP1 sgRNA for 48 hours. WCL was extracted and immunoblot was carried out using the indicated antibodies. (H) Effects of TES1 vs TES2 signaling inhibition on LCL PFKFB4 expression. Median PFKFB4 RNAseq reads from RNAseq analysis of Cas9+ GM12878 expressing LMP1 sgRNA together with dox-induced LMP1 TES1m vs WT LMP1 cDNA expression. Shown are data from n=3 RNAseq datasets. (I) Relative PFKFB4 protein abundances from proteomic analysis of primary human B-cells at the indicated days post-infection by the EBV B95.8 strain. Shown are mean ± SD values from n=4 replicates. (J) Effects of exogenous PFKFB4 expression on proliferation of primary B cells infected by EBV with WT versus TES2m LMP1. Peripheral blood B cells from three independent donors were infected with EBV with WT vs TES2m LMP1. After 14 days, cells were transduced with lentivirus control or with lentivirus driving stable PFKFB4 expression. Five days post-selection, cells were re-seeded. Twelve days later, live cell counts were defined. P-values were determined by one-sided Fisher’s exact test. * p<0.05, **p<0.005, ***p<0.0005. blots are representative of n=3 independent replicates.
    Polya Selection Module, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    polya selected rna  (New England Biolabs)
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    (A) Volcano plot analysis of host transcriptome-wide genes differentially expressed in primary B cells from three independent donors infected with WT or TES2m LMP1 for 21 days. Positive fold changes indicate higher transcripts in cells with TES2m LMP1. (B) PFKFB4 glucose metabolism roles. Glucose is metabolized by glycolysis or by the PPP. Glycolysis flux is regulated via Fructose-2,6-BP, an allosteric regulator of PFK1. PFKFB3 phosphorylates Fructose-6-P into Fructose-2,6-BP while PFKFB4 phosphatase activity converts Fructose-2,6-BP to Fructose-6-P to shunt glycolytic flux to PPP. Created in BioRender. Burton, E. (2026) https://BioRender.com/i0gawc8 . (C) KEGG pathway analysis of transcripts significantly upregulated in cells infected by EBV with WT LMP1 at day 21 post infection. (D) KEGG pathway analysis of transcripts significantly upregulated in cells infected by TES2m EBV at day 21 post infection. (E) PFKFB4 protein levels in WT versus TES2m LMP1 EBV infected cells. Cells from two independent donors were infected with EBV with WT LMP1 or TES2m LMP1 for 22 days. Densitometry values of PFKFB4 vs β-Actin load controls are shown. (F) Effects of LCL LMP1 KO on PFKFB4 <t>mRNA.</t> Median PFKFB4 reads from RNAseq analysis of Cas9+ GM12878 cells with control vs LMP1 targeting sgRNA expression for 48 hours. Shown are data from n=3 RNAseq datasets. (G) PFKFB4 protein levels in LMP1 KO LCLs. Cas9+ GM12878 were mock induced or induced to express LMP1 sgRNA for 48 hours. WCL was extracted and immunoblot was carried out using the indicated antibodies. (H) Effects of TES1 vs TES2 signaling inhibition on LCL PFKFB4 expression. Median PFKFB4 RNAseq reads from RNAseq analysis of Cas9+ GM12878 expressing LMP1 sgRNA together with dox-induced LMP1 TES1m vs WT LMP1 cDNA expression. Shown are data from n=3 RNAseq datasets. (I) Relative PFKFB4 protein abundances from proteomic analysis of primary human B-cells at the indicated days post-infection by the EBV B95.8 strain. Shown are mean ± SD values from n=4 replicates. (J) Effects of exogenous PFKFB4 expression on proliferation of primary B cells infected by EBV with WT versus TES2m LMP1. Peripheral blood B cells from three independent donors were infected with EBV with WT vs TES2m LMP1. After 14 days, cells were transduced with lentivirus control or with lentivirus driving stable PFKFB4 expression. Five days post-selection, cells were re-seeded. Twelve days later, live cell counts were defined. P-values were determined by one-sided Fisher’s exact test. * p<0.05, **p<0.005, ***p<0.0005. blots are representative of n=3 independent replicates.
    Polya Selected Rna, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    (A) Volcano plot analysis of host transcriptome-wide genes differentially expressed in primary B cells from three independent donors infected with WT or TES2m LMP1 for 21 days. Positive fold changes indicate higher transcripts in cells with TES2m LMP1. (B) PFKFB4 glucose metabolism roles. Glucose is metabolized by glycolysis or by the PPP. Glycolysis flux is regulated via Fructose-2,6-BP, an allosteric regulator of PFK1. PFKFB3 phosphorylates Fructose-6-P into Fructose-2,6-BP while PFKFB4 phosphatase activity converts Fructose-2,6-BP to Fructose-6-P to shunt glycolytic flux to PPP. Created in BioRender. Burton, E. (2026) https://BioRender.com/i0gawc8 . (C) KEGG pathway analysis of transcripts significantly upregulated in cells infected by EBV with WT LMP1 at day 21 post infection. (D) KEGG pathway analysis of transcripts significantly upregulated in cells infected by TES2m EBV at day 21 post infection. (E) PFKFB4 protein levels in WT versus TES2m LMP1 EBV infected cells. Cells from two independent donors were infected with EBV with WT LMP1 or TES2m LMP1 for 22 days. Densitometry values of PFKFB4 vs β-Actin load controls are shown. (F) Effects of LCL LMP1 KO on PFKFB4 <t>mRNA.</t> Median PFKFB4 reads from RNAseq analysis of Cas9+ GM12878 cells with control vs LMP1 targeting sgRNA expression for 48 hours. Shown are data from n=3 RNAseq datasets. (G) PFKFB4 protein levels in LMP1 KO LCLs. Cas9+ GM12878 were mock induced or induced to express LMP1 sgRNA for 48 hours. WCL was extracted and immunoblot was carried out using the indicated antibodies. (H) Effects of TES1 vs TES2 signaling inhibition on LCL PFKFB4 expression. Median PFKFB4 RNAseq reads from RNAseq analysis of Cas9+ GM12878 expressing LMP1 sgRNA together with dox-induced LMP1 TES1m vs WT LMP1 cDNA expression. Shown are data from n=3 RNAseq datasets. (I) Relative PFKFB4 protein abundances from proteomic analysis of primary human B-cells at the indicated days post-infection by the EBV B95.8 strain. Shown are mean ± SD values from n=4 replicates. (J) Effects of exogenous PFKFB4 expression on proliferation of primary B cells infected by EBV with WT versus TES2m LMP1. Peripheral blood B cells from three independent donors were infected with EBV with WT vs TES2m LMP1. After 14 days, cells were transduced with lentivirus control or with lentivirus driving stable PFKFB4 expression. Five days post-selection, cells were re-seeded. Twelve days later, live cell counts were defined. P-values were determined by one-sided Fisher’s exact test. * p<0.05, **p<0.005, ***p<0.0005. blots are representative of n=3 independent replicates.
    Polya Selection, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    (A) Volcano plot analysis of host transcriptome-wide genes differentially expressed in primary B cells from three independent donors infected with WT or TES2m LMP1 for 21 days. Positive fold changes indicate higher transcripts in cells with TES2m LMP1. (B) PFKFB4 glucose metabolism roles. Glucose is metabolized by glycolysis or by the PPP. Glycolysis flux is regulated via Fructose-2,6-BP, an allosteric regulator of PFK1. PFKFB3 phosphorylates Fructose-6-P into Fructose-2,6-BP while PFKFB4 phosphatase activity converts Fructose-2,6-BP to Fructose-6-P to shunt glycolytic flux to PPP. Created in BioRender. Burton, E. (2026) https://BioRender.com/i0gawc8 . (C) KEGG pathway analysis of transcripts significantly upregulated in cells infected by EBV with WT LMP1 at day 21 post infection. (D) KEGG pathway analysis of transcripts significantly upregulated in cells infected by TES2m EBV at day 21 post infection. (E) PFKFB4 protein levels in WT versus TES2m LMP1 EBV infected cells. Cells from two independent donors were infected with EBV with WT LMP1 or TES2m LMP1 for 22 days. Densitometry values of PFKFB4 vs β-Actin load controls are shown. (F) Effects of LCL LMP1 KO on PFKFB4 <t>mRNA.</t> Median PFKFB4 reads from RNAseq analysis of Cas9+ GM12878 cells with control vs LMP1 targeting sgRNA expression for 48 hours. Shown are data from n=3 RNAseq datasets. (G) PFKFB4 protein levels in LMP1 KO LCLs. Cas9+ GM12878 were mock induced or induced to express LMP1 sgRNA for 48 hours. WCL was extracted and immunoblot was carried out using the indicated antibodies. (H) Effects of TES1 vs TES2 signaling inhibition on LCL PFKFB4 expression. Median PFKFB4 RNAseq reads from RNAseq analysis of Cas9+ GM12878 expressing LMP1 sgRNA together with dox-induced LMP1 TES1m vs WT LMP1 cDNA expression. Shown are data from n=3 RNAseq datasets. (I) Relative PFKFB4 protein abundances from proteomic analysis of primary human B-cells at the indicated days post-infection by the EBV B95.8 strain. Shown are mean ± SD values from n=4 replicates. (J) Effects of exogenous PFKFB4 expression on proliferation of primary B cells infected by EBV with WT versus TES2m LMP1. Peripheral blood B cells from three independent donors were infected with EBV with WT vs TES2m LMP1. After 14 days, cells were transduced with lentivirus control or with lentivirus driving stable PFKFB4 expression. Five days post-selection, cells were re-seeded. Twelve days later, live cell counts were defined. P-values were determined by one-sided Fisher’s exact test. * p<0.05, **p<0.005, ***p<0.0005. blots are representative of n=3 independent replicates.
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    (A) Volcano plot analysis of host transcriptome-wide genes differentially expressed in primary B cells from three independent donors infected with WT or TES2m LMP1 for 21 days. Positive fold changes indicate higher transcripts in cells with TES2m LMP1. (B) PFKFB4 glucose metabolism roles. Glucose is metabolized by glycolysis or by the PPP. Glycolysis flux is regulated via Fructose-2,6-BP, an allosteric regulator of PFK1. PFKFB3 phosphorylates Fructose-6-P into Fructose-2,6-BP while PFKFB4 phosphatase activity converts Fructose-2,6-BP to Fructose-6-P to shunt glycolytic flux to PPP. Created in BioRender. Burton, E. (2026) https://BioRender.com/i0gawc8 . (C) KEGG pathway analysis of transcripts significantly upregulated in cells infected by EBV with WT LMP1 at day 21 post infection. (D) KEGG pathway analysis of transcripts significantly upregulated in cells infected by TES2m EBV at day 21 post infection. (E) PFKFB4 protein levels in WT versus TES2m LMP1 EBV infected cells. Cells from two independent donors were infected with EBV with WT LMP1 or TES2m LMP1 for 22 days. Densitometry values of PFKFB4 vs β-Actin load controls are shown. (F) Effects of LCL LMP1 KO on PFKFB4 mRNA. Median PFKFB4 reads from RNAseq analysis of Cas9+ GM12878 cells with control vs LMP1 targeting sgRNA expression for 48 hours. Shown are data from n=3 RNAseq datasets. (G) PFKFB4 protein levels in LMP1 KO LCLs. Cas9+ GM12878 were mock induced or induced to express LMP1 sgRNA for 48 hours. WCL was extracted and immunoblot was carried out using the indicated antibodies. (H) Effects of TES1 vs TES2 signaling inhibition on LCL PFKFB4 expression. Median PFKFB4 RNAseq reads from RNAseq analysis of Cas9+ GM12878 expressing LMP1 sgRNA together with dox-induced LMP1 TES1m vs WT LMP1 cDNA expression. Shown are data from n=3 RNAseq datasets. (I) Relative PFKFB4 protein abundances from proteomic analysis of primary human B-cells at the indicated days post-infection by the EBV B95.8 strain. Shown are mean ± SD values from n=4 replicates. (J) Effects of exogenous PFKFB4 expression on proliferation of primary B cells infected by EBV with WT versus TES2m LMP1. Peripheral blood B cells from three independent donors were infected with EBV with WT vs TES2m LMP1. After 14 days, cells were transduced with lentivirus control or with lentivirus driving stable PFKFB4 expression. Five days post-selection, cells were re-seeded. Twelve days later, live cell counts were defined. P-values were determined by one-sided Fisher’s exact test. * p<0.05, **p<0.005, ***p<0.0005. blots are representative of n=3 independent replicates.

    Journal: bioRxiv

    Article Title: Epstein-Barr Virus Latent Membrane Protein 1 Suppresses Ferroptosis via Pentose Phosphate Pathway and Glutathione Metabolism

    doi: 10.64898/2026.03.09.710628

    Figure Lengend Snippet: (A) Volcano plot analysis of host transcriptome-wide genes differentially expressed in primary B cells from three independent donors infected with WT or TES2m LMP1 for 21 days. Positive fold changes indicate higher transcripts in cells with TES2m LMP1. (B) PFKFB4 glucose metabolism roles. Glucose is metabolized by glycolysis or by the PPP. Glycolysis flux is regulated via Fructose-2,6-BP, an allosteric regulator of PFK1. PFKFB3 phosphorylates Fructose-6-P into Fructose-2,6-BP while PFKFB4 phosphatase activity converts Fructose-2,6-BP to Fructose-6-P to shunt glycolytic flux to PPP. Created in BioRender. Burton, E. (2026) https://BioRender.com/i0gawc8 . (C) KEGG pathway analysis of transcripts significantly upregulated in cells infected by EBV with WT LMP1 at day 21 post infection. (D) KEGG pathway analysis of transcripts significantly upregulated in cells infected by TES2m EBV at day 21 post infection. (E) PFKFB4 protein levels in WT versus TES2m LMP1 EBV infected cells. Cells from two independent donors were infected with EBV with WT LMP1 or TES2m LMP1 for 22 days. Densitometry values of PFKFB4 vs β-Actin load controls are shown. (F) Effects of LCL LMP1 KO on PFKFB4 mRNA. Median PFKFB4 reads from RNAseq analysis of Cas9+ GM12878 cells with control vs LMP1 targeting sgRNA expression for 48 hours. Shown are data from n=3 RNAseq datasets. (G) PFKFB4 protein levels in LMP1 KO LCLs. Cas9+ GM12878 were mock induced or induced to express LMP1 sgRNA for 48 hours. WCL was extracted and immunoblot was carried out using the indicated antibodies. (H) Effects of TES1 vs TES2 signaling inhibition on LCL PFKFB4 expression. Median PFKFB4 RNAseq reads from RNAseq analysis of Cas9+ GM12878 expressing LMP1 sgRNA together with dox-induced LMP1 TES1m vs WT LMP1 cDNA expression. Shown are data from n=3 RNAseq datasets. (I) Relative PFKFB4 protein abundances from proteomic analysis of primary human B-cells at the indicated days post-infection by the EBV B95.8 strain. Shown are mean ± SD values from n=4 replicates. (J) Effects of exogenous PFKFB4 expression on proliferation of primary B cells infected by EBV with WT versus TES2m LMP1. Peripheral blood B cells from three independent donors were infected with EBV with WT vs TES2m LMP1. After 14 days, cells were transduced with lentivirus control or with lentivirus driving stable PFKFB4 expression. Five days post-selection, cells were re-seeded. Twelve days later, live cell counts were defined. P-values were determined by one-sided Fisher’s exact test. * p<0.05, **p<0.005, ***p<0.0005. blots are representative of n=3 independent replicates.

    Article Snippet: To construct indexed libraries, 1 μg of total RNA was used for polyA mRNA selection using NEBNext Poly(A) mRNA Magnetic Isolation Module (Cat#E7490S), and library preparation with NEBNext Ultra RNA Library Prep with Sample Purification Beads (Cat#E7765S).

    Techniques: Infection, Activity Assay, RNA sequencing, Control, Expressing, Western Blot, Inhibition, Transduction, Selection