normal lung fibroblast cell line mrc  (ATCC)

Journal: International Journal of Molecular Sciences

Article Title: Human Small Airway Epithelia Reveal Dichloroacetate as a Broad-Spectrum Antiviral Against Respiratory Viruses

doi: 10.3390/ijms26209853

Figure Lengend Snippet: Coronavirus infection alters the energy metabolism of MRC5 cells. ( A , B ) Representative images of MRC5 cells cultured in an XFp microwell plate and infected with the coronavirus HCoV-229E. The arrows point to the circular well bumps at the bottom of the plate that define the height of the microchamber. ( A ) Bright field microscopy image of MRC5 cells seeded in an XFp microwell. ( B ) Fluorescence microscopy image showing cells infected with the GFP-labeled virus HCoV-229E in the XFp well. ( C – E ) Effect of viral infection on cell bioenergetics. ( C ) Rate of respiration (OCR), ( D ) aerobic glycolysis (ECAR), and ( E ) metabolic profile (the OCR/ECAR ratio). The data are presented as the mean ± s.e.m. of 4 biologically independent experiments performed in triplicate. The statistical significance of the differences was calculated via an unpaired Student’s t test. * p < 0.05; *** p < 0.001.

Article Snippet: The human cell lines MRC5 (lung fibroblasts), hSAEC (primary small airway epithelial cells), Huh7 (hepatoma cancer cells) and Hep2 (carcinoma) were obtained from the American Type Culture Collection (Life Technologies, Frederick, MD, USA).

Techniques: Infection, Cell Culture, Microscopy, Fluorescence, Labeling, Virus

Journal: International Journal of Molecular Sciences

Article Title: Human Small Airway Epithelia Reveal Dichloroacetate as a Broad-Spectrum Antiviral Against Respiratory Viruses

doi: 10.3390/ijms26209853

Figure Lengend Snippet: The antiviral activity of PI3K/AKT/mTOR signaling pathway inhibitors in MRC5 cells infected with HCoV-229E. ( A ) Scheme of the signaling pathway and the inhibitors whose antiviral activity was studied. Effect of the pathway inhibitors on the progression of infection in the lung fibroblast cell line MRC5 ( B – D ) or the hepatoma cell line Huh7 ( E – G ); ( B , E ) percentage of infected cells; ( C , F ) mean GFP fluorescence of the infected cells; ( D , G ) overall virus gene expression calculated from the percentage of infected cells and the mean fluorescence intensity. The bars represent the mean ± s.e.m. values, and the number of biologically independent experiments ranged from 6 to 12 (MRC5) or 5 to 7 (Huh7); all the experiments were performed in triplicate. The statistical significance of the differences was calculated using an unpaired Student’s t test relative to the untreated control. * p < 0.05; ** p < 0.01; *** p < 0.001. Where not indicated, the differences were not significant. Abbreviations: PM, plasma membrane; MMs, mitochondrial membranes; LPT, lapatinib; WTM, wortmaninn; EVL, everolimus; 2ME, 2-methoxyestradiol; DCA, dichloroacetate.

Article Snippet: The human cell lines MRC5 (lung fibroblasts), hSAEC (primary small airway epithelial cells), Huh7 (hepatoma cancer cells) and Hep2 (carcinoma) were obtained from the American Type Culture Collection (Life Technologies, Frederick, MD, USA).

Techniques: Activity Assay, Infection, Fluorescence, Virus, Gene Expression, Control, Clinical Proteomics, Membrane

Journal: International Journal of Molecular Sciences

Article Title: Human Small Airway Epithelia Reveal Dichloroacetate as a Broad-Spectrum Antiviral Against Respiratory Viruses

doi: 10.3390/ijms26209853

Figure Lengend Snippet: Inhibitors of the PI3K/AKT/mTOR signaling pathway differentially alter the metabolism of uninfected MRC5 and Huh7 cells. The effect of PI3K/AKT/mTOR pathway inhibitors on the energy metabolism of MRC5 ( A – C ) and Huh7 ( D – F ) cells: ( A , D ) Rate of respiration (OCR), ( B , E ) aerobic glycolysis (ECAR), and ( C , F ) metabolic profile (OCR/ECAR ratio). The data are presented as the mean ± s.e.m. of 5–14 biologically independent experiments performed with 5 technical replicates. The statistical significance of the differences was calculated using an unpaired Student’s t test relative to the untreated control. ** p < 0.01; *** p < 0.001. Where not indicated, the differences were not significant. Abbreviations: LPT, lapatinib; WTM, wortmaninn; EVL, everolimus; 2ME, 2-methoxyestradiol; DCA, dichloroacetate. Dashed line indicates control value in every case.

Article Snippet: The human cell lines MRC5 (lung fibroblasts), hSAEC (primary small airway epithelial cells), Huh7 (hepatoma cancer cells) and Hep2 (carcinoma) were obtained from the American Type Culture Collection (Life Technologies, Frederick, MD, USA).

Techniques: Control