mrc5 human embryonic lung fibroblast cells  (ATCC)

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: 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 .

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

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) Sequence alignment of hsa_circCPSF6 and mmu_circCpsf6 was performed using basic local alignment search tool (BLAST). (B - C) CircCPSF6 expression level determined by RT-PCR in NDV infected (MOI 1; 24h) (B) A549 cells and (C) MRC5 cells.

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 .

Techniques: Sequencing, Expressing, Reverse Transcription Polymerase Chain Reaction, Infection

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) CircCPSF6 knockdown and viral load was measured by RT-PCR in PR8 infected (MOI 1; 24h) MRC5 cells. (B) Knockdown of circCPSF6 measured by RT-PCR in PR8 infected (MOI 1; 24h) human PBMCs, mice mpLFs and BMDCs. (C – D) Viral load measured by RT-PCR in circCPSF6 knockdown A549 cells followed by (C) NDV infection (MOI 1; 24h) and (D) SeV infection (MOI 2; 24h). (E) Overexpression level of circCPSF6 and viral load measured by RT-PCR in PR8 infected (MOI 1; 24h) MRC5 cells. (F) Viral load measured by RT-PCR in circCPSF6 overexpressing, NDV infected (MOI 1; 24h) A549 cells. (G) IL-6 transcript expression measured by RT-PCR in circCPSF6 knockdown, PR8 infected A549 cells (MOI 1; 24h).

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 .

Techniques: Knockdown, Reverse Transcription Polymerase Chain Reaction, Infection, Over Expression, Expressing

Journal: Journal of Clinical Microbiology

Article Title: High-Content Screening, a Reliable System for Coxiella burnetii Isolation from Clinical Samples

doi: 10.1128/JCM.02081-19

Figure Lengend Snippet: Traditional and improved coculture and detection processes adopted for the isolation of C. burnetii. This figure details all the steps involved in processing a clinical sample using conventional shell vial analysis and the new optimized high-content screening technique.

Article Snippet: Two cell lines were used as cellular supports for coculture: human embryonic lung fibroblast MRC5 cells (RD-Biotech, Besançon, France) and mouse fibroblast L929 cells (ATCC CCL-1).

Techniques: Isolation, High Content Screening

Journal: Journal of Clinical Microbiology

Article Title: High-Content Screening, a Reliable System for Coxiella burnetii Isolation from Clinical Samples

doi: 10.1128/JCM.02081-19

Figure Lengend Snippet: Comparative results between the shell vial assay and the HCS assay for the isolation of C. burnetii from clinical samples. This interaction scheme shows all positive samples isolated using the two strategies, the samples’ origins (nodes), and their coculture delay (lines).

Article Snippet: Two cell lines were used as cellular supports for coculture: human embryonic lung fibroblast MRC5 cells (RD-Biotech, Besançon, France) and mouse fibroblast L929 cells (ATCC CCL-1).

Techniques: Isolation

Journal: bioRxiv

Article Title: High Content Screening, a reliable system for Coxiella burnetii isolation from clinical samples

doi: 10.1101/2019.12.17.880484

Figure Lengend Snippet: Cell density of L929 and MRC5 cell lines at different initial concentrations, 24 h into culture. (a), (b), (c) and (d) represent respective brightfield images of L929 cells at 10 6 , 4×10 5 , 2×10 5 and 10 5 cells/ml. (e), (f), (g) and (h) represent respective concentrations for MRC5 cells. Scale bars indicate 100 µm.

Article Snippet: Two cell lines were used as cellular supports for co-culture: the human embryonic lung fibroblast MRC5 cells (RD-Biotech, Besançon, France) and the mouse fibroblast L929 cells (ATCC® CCL-1).

Techniques:

Journal: bioRxiv

Article Title: High Content Screening, a reliable system for Coxiella burnetii isolation from clinical samples

doi: 10.1101/2019.12.17.880484

Figure Lengend Snippet: Results from the prediction algorithm and validation references of MRC5 cells infected with CB NMII at 10, 20 and 30 days post infection. (A): The heat map represents the percentages of infected cells obtained with the prediction algorithm of MRC5 cells infected with serial dilutions of CB NMII. (C): The table represents the manual quantification results from the same experiment. (B): Respective immunofluorescence images of (a) the negative control (well B1 in the heat map), (b) the false positive result (well D9), (c) a positive result (well B3) and (d) a false negative result (well B5). (D): Respective fluorescence and brightfield images of (e, f) the negative control, (g, h) cells at an advanced stage of infection and (i, j) slightly infected cells (vacuoles are indicated with red arrows). Scale bars represent 100 µm.

Article Snippet: Two cell lines were used as cellular supports for co-culture: the human embryonic lung fibroblast MRC5 cells (RD-Biotech, Besançon, France) and the mouse fibroblast L929 cells (ATCC® CCL-1).

Techniques: Infection, Immunofluorescence, Negative Control, Fluorescence