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thp1 human peripheral blood monocytes  (ATCC)


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    ATCC thp1 human peripheral blood monocytes
    (A) Heatmap showing clustering of log2 transformed reads per million (RPM) value of circRNAs from RNA seq data (GSE172315) in IAV H9N2 infected A549 cells with relative expression values indicated by blue to red scale (blue, high; red, low). (B) CircCPSF6 expression in control and infected sample from same data. (C) RT-PCR validation of circCPSF6 expression level in A549 cells, infected with PR8 at indicated multiplicity of infection (MOI) and post infection time points. (D) PCR amplification of circCPSF6 and linear CPSF6 using divergent (circRNA specific) and convergent (linear RNA) primers in cDNA or genomic DNA (gDNA) from A549 cells; GAPDH used as control. (E) Schematic representation of circCPSF6 genomic location generated by the back splicing of CPSF6 exons and Sanger sequencing of divergent primer amplicon showing back splice junction (BSJ). (F) Expression of circCPSF6, linear CPSF6 and GAPDH measured with or without RNase R (2U/μg RNA) treatment at 37℃ for 20 mins in A549 cells. (G) Expression of circCPSF6 and linear CPSF6 measured in actinomycin D (2μg/mL) treated A549 cells for indicated time. (H) CircCPSF6 amplification from cDNA prepared using random hexamer and oligo-dT primer. (I) CircCPSF6 copy number quantified by digital PCR (dPCR) in PR8 infected (MOI 1; 24h) A549 cells. (J – L) CircCPSF6 expression in human cell lines (J) MRC5 and <t>THP1,</t> (K) human primary PBMCs, (L) murine primary lung fibroblasts (mpLF) and BMDCs, post PR8 infection (MOI 1; 24h). (M) Schematic representation of in vivo virus challenge and expression of circCpsf6 in mice lung tissue after PR8 infection (PFU 100; 3 days). (N) RNA-FISH using Cy3 labelled antisense-probe designed specific to circCPSF6 BSJ (red) for the cellular localization. Nuclei were stained with DAPI (blue). Scale bar, 10μm. Data are presented as the mean ± SEM from triplicate samples of a single experiment and representative of three independent experiments. **** p < 0.0001, *** p < 0.001, ** p < 0.01 by two-tailed unpaired Student’s t -test or two-way ANOVA test. (Schematics are created with BioRender.com)
    Thp1 Human Peripheral Blood Monocytes, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 20220 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/thp1+human+peripheral+blood+monocytes/THP-1/bio_rxiv__2025__11__03__686289-43-0-5
    Average 99 stars, based on 20220 article reviews
    thp1 human peripheral blood monocytes - by Bioz Stars, 2026-09
    99/100 stars

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    1) Product Images from "A novel role of circCPSF6 regulating antiviral innate immunity via miR-665 and PCBP2-IPS-1 axis"

    Article Title: A novel role of circCPSF6 regulating antiviral innate immunity via miR-665 and PCBP2-IPS-1 axis

    Journal: bioRxiv

    doi: 10.1101/2025.11.03.686289

    (A) Heatmap showing clustering of log2 transformed reads per million (RPM) value of circRNAs from RNA seq data (GSE172315) in IAV H9N2 infected A549 cells with relative expression values indicated by blue to red scale (blue, high; red, low). (B) CircCPSF6 expression in control and infected sample from same data. (C) RT-PCR validation of circCPSF6 expression level in A549 cells, infected with PR8 at indicated multiplicity of infection (MOI) and post infection time points. (D) PCR amplification of circCPSF6 and linear CPSF6 using divergent (circRNA specific) and convergent (linear RNA) primers in cDNA or genomic DNA (gDNA) from A549 cells; GAPDH used as control. (E) Schematic representation of circCPSF6 genomic location generated by the back splicing of CPSF6 exons and Sanger sequencing of divergent primer amplicon showing back splice junction (BSJ). (F) Expression of circCPSF6, linear CPSF6 and GAPDH measured with or without RNase R (2U/μg RNA) treatment at 37℃ for 20 mins in A549 cells. (G) Expression of circCPSF6 and linear CPSF6 measured in actinomycin D (2μg/mL) treated A549 cells for indicated time. (H) CircCPSF6 amplification from cDNA prepared using random hexamer and oligo-dT primer. (I) CircCPSF6 copy number quantified by digital PCR (dPCR) in PR8 infected (MOI 1; 24h) A549 cells. (J – L) CircCPSF6 expression in human cell lines (J) MRC5 and THP1, (K) human primary PBMCs, (L) murine primary lung fibroblasts (mpLF) and BMDCs, post PR8 infection (MOI 1; 24h). (M) Schematic representation of in vivo virus challenge and expression of circCpsf6 in mice lung tissue after PR8 infection (PFU 100; 3 days). (N) RNA-FISH using Cy3 labelled antisense-probe designed specific to circCPSF6 BSJ (red) for the cellular localization. Nuclei were stained with DAPI (blue). Scale bar, 10μm. Data are presented as the mean ± SEM from triplicate samples of a single experiment and representative of three independent experiments. **** p < 0.0001, *** p < 0.001, ** p < 0.01 by two-tailed unpaired Student’s t -test or two-way ANOVA test. (Schematics are created with BioRender.com)
    Figure Legend Snippet: (A) Heatmap showing clustering of log2 transformed reads per million (RPM) value of circRNAs from RNA seq data (GSE172315) in IAV H9N2 infected A549 cells with relative expression values indicated by blue to red scale (blue, high; red, low). (B) CircCPSF6 expression in control and infected sample from same data. (C) RT-PCR validation of circCPSF6 expression level in A549 cells, infected with PR8 at indicated multiplicity of infection (MOI) and post infection time points. (D) PCR amplification of circCPSF6 and linear CPSF6 using divergent (circRNA specific) and convergent (linear RNA) primers in cDNA or genomic DNA (gDNA) from A549 cells; GAPDH used as control. (E) Schematic representation of circCPSF6 genomic location generated by the back splicing of CPSF6 exons and Sanger sequencing of divergent primer amplicon showing back splice junction (BSJ). (F) Expression of circCPSF6, linear CPSF6 and GAPDH measured with or without RNase R (2U/μg RNA) treatment at 37℃ for 20 mins in A549 cells. (G) Expression of circCPSF6 and linear CPSF6 measured in actinomycin D (2μg/mL) treated A549 cells for indicated time. (H) CircCPSF6 amplification from cDNA prepared using random hexamer and oligo-dT primer. (I) CircCPSF6 copy number quantified by digital PCR (dPCR) in PR8 infected (MOI 1; 24h) A549 cells. (J – L) CircCPSF6 expression in human cell lines (J) MRC5 and THP1, (K) human primary PBMCs, (L) murine primary lung fibroblasts (mpLF) and BMDCs, post PR8 infection (MOI 1; 24h). (M) Schematic representation of in vivo virus challenge and expression of circCpsf6 in mice lung tissue after PR8 infection (PFU 100; 3 days). (N) RNA-FISH using Cy3 labelled antisense-probe designed specific to circCPSF6 BSJ (red) for the cellular localization. Nuclei were stained with DAPI (blue). Scale bar, 10μm. Data are presented as the mean ± SEM from triplicate samples of a single experiment and representative of three independent experiments. **** p < 0.0001, *** p < 0.001, ** p < 0.01 by two-tailed unpaired Student’s t -test or two-way ANOVA test. (Schematics are created with BioRender.com)

    Techniques Used: Transformation Assay, RNA Sequencing, Infection, Expressing, Control, Reverse Transcription Polymerase Chain Reaction, Biomarker Discovery, Amplification, Generated, Sequencing, Random Hexamer, Digital PCR, In Vivo, Virus, Staining, Two Tailed Test

    Related Articles

    Transformation Assay:

    Article Title: A novel role of circCPSF6 regulating antiviral innate immunity via miR-665 and PCBP2-IPS-1 axis
    Article Snippet: A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .. THP1 human peripheral blood monocytes (ATCC TIB-202) were maintained in RPMI-1640 Medium supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .

    RNA Sequencing:

    Article Title: A novel role of circCPSF6 regulating antiviral innate immunity via miR-665 and PCBP2-IPS-1 axis
    Article Snippet: A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .. THP1 human peripheral blood monocytes (ATCC TIB-202) were maintained in RPMI-1640 Medium supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .

    Infection:

    Article Title: A novel role of circCPSF6 regulating antiviral innate immunity via miR-665 and PCBP2-IPS-1 axis
    Article Snippet: A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .. THP1 human peripheral blood monocytes (ATCC TIB-202) were maintained in RPMI-1640 Medium supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .

    Expressing:

    Article Title: A novel role of circCPSF6 regulating antiviral innate immunity via miR-665 and PCBP2-IPS-1 axis
    Article Snippet: A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .. THP1 human peripheral blood monocytes (ATCC TIB-202) were maintained in RPMI-1640 Medium supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .

    Control:

    Article Title: A novel role of circCPSF6 regulating antiviral innate immunity via miR-665 and PCBP2-IPS-1 axis
    Article Snippet: A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .. THP1 human peripheral blood monocytes (ATCC TIB-202) were maintained in RPMI-1640 Medium supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: A novel role of circCPSF6 regulating antiviral innate immunity via miR-665 and PCBP2-IPS-1 axis
    Article Snippet: A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .. THP1 human peripheral blood monocytes (ATCC TIB-202) were maintained in RPMI-1640 Medium supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .

    Biomarker Discovery:

    Article Title: A novel role of circCPSF6 regulating antiviral innate immunity via miR-665 and PCBP2-IPS-1 axis
    Article Snippet: A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .. THP1 human peripheral blood monocytes (ATCC TIB-202) were maintained in RPMI-1640 Medium supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .

    Amplification:

    Article Title: A novel role of circCPSF6 regulating antiviral innate immunity via miR-665 and PCBP2-IPS-1 axis
    Article Snippet: A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .. THP1 human peripheral blood monocytes (ATCC TIB-202) were maintained in RPMI-1640 Medium supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .

    Generated:

    Article Title: A novel role of circCPSF6 regulating antiviral innate immunity via miR-665 and PCBP2-IPS-1 axis
    Article Snippet: A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .. THP1 human peripheral blood monocytes (ATCC TIB-202) were maintained in RPMI-1640 Medium supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .

    Sequencing:

    Article Title: A novel role of circCPSF6 regulating antiviral innate immunity via miR-665 and PCBP2-IPS-1 axis
    Article Snippet: A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .. THP1 human peripheral blood monocytes (ATCC TIB-202) were maintained in RPMI-1640 Medium supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .

    Random Hexamer:

    Article Title: A novel role of circCPSF6 regulating antiviral innate immunity via miR-665 and PCBP2-IPS-1 axis
    Article Snippet: A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .. THP1 human peripheral blood monocytes (ATCC TIB-202) were maintained in RPMI-1640 Medium supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .

    Digital PCR:

    Article Title: A novel role of circCPSF6 regulating antiviral innate immunity via miR-665 and PCBP2-IPS-1 axis
    Article Snippet: A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .. THP1 human peripheral blood monocytes (ATCC TIB-202) were maintained in RPMI-1640 Medium supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .

    In Vivo:

    Article Title: A novel role of circCPSF6 regulating antiviral innate immunity via miR-665 and PCBP2-IPS-1 axis
    Article Snippet: A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .. THP1 human peripheral blood monocytes (ATCC TIB-202) were maintained in RPMI-1640 Medium supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .

    Virus:

    Article Title: A novel role of circCPSF6 regulating antiviral innate immunity via miR-665 and PCBP2-IPS-1 axis
    Article Snippet: A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .. THP1 human peripheral blood monocytes (ATCC TIB-202) were maintained in RPMI-1640 Medium supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .

    Staining:

    Article Title: A novel role of circCPSF6 regulating antiviral innate immunity via miR-665 and PCBP2-IPS-1 axis
    Article Snippet: A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .. THP1 human peripheral blood monocytes (ATCC TIB-202) were maintained in RPMI-1640 Medium supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .

    Two Tailed Test:

    Article Title: A novel role of circCPSF6 regulating antiviral innate immunity via miR-665 and PCBP2-IPS-1 axis
    Article Snippet: A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .A549 human alveolar epithelial cells (ATCC CCL-185), HEK293T human embryonic kidney cells (ATCC CRL-3216), MRC5 human embryonic lung fibroblast cells (ATCC CCL-171) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .. THP1 human peripheral blood monocytes (ATCC TIB-202) were maintained in RPMI-1640 Medium supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .



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    ATCC human peripheral blood monocytic thp1 cells
    (A) Heatmap showing clustering of log2 transformed reads per million (RPM) value of circRNAs from RNA seq data (GSE172315) in IAV H9N2 infected A549 cells with relative expression values indicated by blue to red scale (blue, high; red, low). (B) CircCPSF6 expression in control and infected sample from same data. (C) RT-PCR validation of circCPSF6 expression level in A549 cells, infected with PR8 at indicated multiplicity of infection (MOI) and post infection time points. (D) PCR amplification of circCPSF6 and linear CPSF6 using divergent (circRNA specific) and convergent (linear RNA) primers in cDNA or genomic DNA (gDNA) from A549 cells; GAPDH used as control. (E) Schematic representation of circCPSF6 genomic location generated by the back splicing of CPSF6 exons and Sanger sequencing of divergent primer amplicon showing back splice junction (BSJ). (F) Expression of circCPSF6, linear CPSF6 and GAPDH measured with or without RNase R (2U/μg RNA) treatment at 37℃ for 20 mins in A549 cells. (G) Expression of circCPSF6 and linear CPSF6 measured in actinomycin D (2μg/mL) treated A549 cells for indicated time. (H) CircCPSF6 amplification from cDNA prepared using random hexamer and oligo-dT primer. (I) CircCPSF6 copy number quantified by digital PCR (dPCR) in PR8 infected (MOI 1; 24h) A549 cells. (J – L) CircCPSF6 expression in human cell lines (J) MRC5 and <t>THP1,</t> (K) human primary PBMCs, (L) murine primary lung fibroblasts (mpLF) and BMDCs, post PR8 infection (MOI 1; 24h). (M) Schematic representation of in vivo virus challenge and expression of circCpsf6 in mice lung tissue after PR8 infection (PFU 100; 3 days). (N) RNA-FISH using Cy3 labelled antisense-probe designed specific to circCPSF6 BSJ (red) for the cellular localization. Nuclei were stained with DAPI (blue). Scale bar, 10μm. Data are presented as the mean ± SEM from triplicate samples of a single experiment and representative of three independent experiments. **** p < 0.0001, *** p < 0.001, ** p < 0.01 by two-tailed unpaired Student’s t -test or two-way ANOVA test. (Schematics are created with BioRender.com)
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    (A) Heatmap showing clustering of log2 transformed reads per million (RPM) value of circRNAs from RNA seq data (GSE172315) in IAV H9N2 infected A549 cells with relative expression values indicated by blue to red scale (blue, high; red, low). (B) CircCPSF6 expression in control and infected sample from same data. (C) RT-PCR validation of circCPSF6 expression level in A549 cells, infected with PR8 at indicated multiplicity of infection (MOI) and post infection time points. (D) PCR amplification of circCPSF6 and linear CPSF6 using divergent (circRNA specific) and convergent (linear RNA) primers in cDNA or genomic DNA (gDNA) from A549 cells; GAPDH used as control. (E) Schematic representation of circCPSF6 genomic location generated by the back splicing of CPSF6 exons and Sanger sequencing of divergent primer amplicon showing back splice junction (BSJ). (F) Expression of circCPSF6, linear CPSF6 and GAPDH measured with or without RNase R (2U/μg RNA) treatment at 37℃ for 20 mins in A549 cells. (G) Expression of circCPSF6 and linear CPSF6 measured in actinomycin D (2μg/mL) treated A549 cells for indicated time. (H) CircCPSF6 amplification from cDNA prepared using random hexamer and oligo-dT primer. (I) CircCPSF6 copy number quantified by digital PCR (dPCR) in PR8 infected (MOI 1; 24h) A549 cells. (J – L) CircCPSF6 expression in human cell lines (J) MRC5 and THP1, (K) human primary PBMCs, (L) murine primary lung fibroblasts (mpLF) and BMDCs, post PR8 infection (MOI 1; 24h). (M) Schematic representation of in vivo virus challenge and expression of circCpsf6 in mice lung tissue after PR8 infection (PFU 100; 3 days). (N) RNA-FISH using Cy3 labelled antisense-probe designed specific to circCPSF6 BSJ (red) for the cellular localization. Nuclei were stained with DAPI (blue). Scale bar, 10μm. Data are presented as the mean ± SEM from triplicate samples of a single experiment and representative of three independent experiments. **** p < 0.0001, *** p < 0.001, ** p < 0.01 by two-tailed unpaired Student’s t -test or two-way ANOVA test. (Schematics are created with BioRender.com)

    Journal: bioRxiv

    Article Title: A novel role of circCPSF6 regulating antiviral innate immunity via miR-665 and PCBP2-IPS-1 axis

    doi: 10.1101/2025.11.03.686289

    Figure Lengend Snippet: (A) Heatmap showing clustering of log2 transformed reads per million (RPM) value of circRNAs from RNA seq data (GSE172315) in IAV H9N2 infected A549 cells with relative expression values indicated by blue to red scale (blue, high; red, low). (B) CircCPSF6 expression in control and infected sample from same data. (C) RT-PCR validation of circCPSF6 expression level in A549 cells, infected with PR8 at indicated multiplicity of infection (MOI) and post infection time points. (D) PCR amplification of circCPSF6 and linear CPSF6 using divergent (circRNA specific) and convergent (linear RNA) primers in cDNA or genomic DNA (gDNA) from A549 cells; GAPDH used as control. (E) Schematic representation of circCPSF6 genomic location generated by the back splicing of CPSF6 exons and Sanger sequencing of divergent primer amplicon showing back splice junction (BSJ). (F) Expression of circCPSF6, linear CPSF6 and GAPDH measured with or without RNase R (2U/μg RNA) treatment at 37℃ for 20 mins in A549 cells. (G) Expression of circCPSF6 and linear CPSF6 measured in actinomycin D (2μg/mL) treated A549 cells for indicated time. (H) CircCPSF6 amplification from cDNA prepared using random hexamer and oligo-dT primer. (I) CircCPSF6 copy number quantified by digital PCR (dPCR) in PR8 infected (MOI 1; 24h) A549 cells. (J – L) CircCPSF6 expression in human cell lines (J) MRC5 and THP1, (K) human primary PBMCs, (L) murine primary lung fibroblasts (mpLF) and BMDCs, post PR8 infection (MOI 1; 24h). (M) Schematic representation of in vivo virus challenge and expression of circCpsf6 in mice lung tissue after PR8 infection (PFU 100; 3 days). (N) RNA-FISH using Cy3 labelled antisense-probe designed specific to circCPSF6 BSJ (red) for the cellular localization. Nuclei were stained with DAPI (blue). Scale bar, 10μm. Data are presented as the mean ± SEM from triplicate samples of a single experiment and representative of three independent experiments. **** p < 0.0001, *** p < 0.001, ** p < 0.01 by two-tailed unpaired Student’s t -test or two-way ANOVA test. (Schematics are created with BioRender.com)

    Article Snippet: THP1 human peripheral blood monocytes (ATCC TIB-202) were maintained in RPMI-1640 Medium supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37°C in 5% CO 2 .

    Techniques: Transformation Assay, RNA Sequencing, Infection, Expressing, Control, Reverse Transcription Polymerase Chain Reaction, Biomarker Discovery, Amplification, Generated, Sequencing, Random Hexamer, Digital PCR, In Vivo, Virus, Staining, Two Tailed Test