Journal: Frontiers in Pharmacology

Article Title: A Herbal Mixture Formula of OCD20015-V009 Prophylactic Administration to Enhance Interferon-Mediated Antiviral Activity Against Influenza A Virus

doi: 10.3389/fphar.2021.764297

Figure Lengend Snippet: The phytochemical profile of OCD20015-V009 by UHPLC-MS/MS.

Article Snippet: Ajugol, calycosin, calycosin-7-glucoside, catalpol, chlorogenic acid, epicatechin, formononetin, ginsenoside Rb1, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rf, ginsenoside Rg1, ginsenoside Rg5, ginsenoside Rk2, ginsenoside Ro, isoquercitrin, loganic acid, and notoginsenoside R2 (Targetmol, United States) were purchased.

Techniques:

Journal: Nature Communications

Article Title: The arginine methyltransferase PRMT7 promotes extravasation of monocytes resulting in tissue injury in COPD

doi: 10.1038/s41467-022-28809-4

Figure Lengend Snippet: a Heat map of normalized enrichment score (NES) following GSEA of cell death pathway gene lists on array data from lung tissue of COPD patients (GSE47460-GPL14550; 145 COPD patients vs 91 healthy controls). b Expression of ferroptotic genes in the lungs of COPD patients relative to healthy controls taken from GSE47460-GPL14550, relative fold change and P value calculated using the GEO2R interactive web tool running limma R . c Western blot analysis of ACSL4 expression in lung core biopsies from healthy ( n = 3) and COPD patients ( n = 3), quantification relative to vinculin shown for individual patients. d Representative images of immunohistochemical analysis for ACSL4 (brown signal indicated by arrowhead, hematoxylin counterstained, scale bar 25 μm) in lung sections from COPD patients ( n = 4) and healthy controls ( n = 4). e Quantification of alveolar epithelial cells positive for ACSL4. f Heat map of NES following GSEA of cell death pathway gene lists on array data from mice exposed to 4 m chronic cigarette smoke (CS, n = 3) v filtered air (FA, n = 3). g Expression of ferroptotic genes in the lungs of mice exposed to 4 m chronic CS ( n = 3) relative to FA ( n = 3) taken from array data used in ( f ), relative fold change and P value calculated using the GEO2R interactive web tool running limma R. h , i Cells from whole lung suspensions of mice exposed to FA ( n = 3) or CS for 4 m ( n = 5), were analysed by scRNA-Seq (Drop-Seq). h UMAP of scRNA-Seq profiles (dots) colored by cell type. i UMAP plots showing expression of ACSL4 in scRNA-Seq profiles. j Representative images of immunohistochemical analysis for ACSL4 (red signal indicated by arrowheads, hematoxylin counterstained, scale bar 25 μm) in lung sections from WT and Prmt7 +/− mice exposed to FA or CS for 4 months ( n = 4–5 per group). k Quantification of alveolar epithelial cells positive for ACSL4 from ( j ) ( n = 5 for WT FA, Prmt7 +/ − FA, Prmt7 +/− CS and n = 4 for WT CS). l , m Mouse AT2 cells (MLE12) were treated with RSL3 (250 nM), or conditioned medium from control macrophages (MФ, RAW264.7 cells) or those polarized to a pro-inflammatory (IFNγ + LPS) or anti-inflammatory (IL4) phenotype for 48 h, representative data shown from a single experiment repeated twice. l Representative image showing PI uptake at 24 h (Scale bar 50 µm). m Number of dead AT2 cells at the time indicated, data shown from a single experiment repeated twice. n BODIPY oxidation in AT2 cells (MLE12) at 6 h after treatment with RSL3 (250 nM, n = 3) or conditioned medium from control macrophages ( n = 2) or those polarized to a pro-inflammatory ( n = 6) or anti-inflammatory ( n = 3) phenotype . o Number of dead AT2 cells (MLE12) by PI uptake at the time indicated following treatment with RSL3 (250 nM), or conditioned medium from macrophages (MФ, RAW264.7 cells) polarized to pro-inflammatory (IFNγ + LPS) plus 20 μM zVAD or 10 μM necrostatin-1s (Nec1-s), representative data shown from a single experiment repeated twice. p Western blot analysis of ACSL4 in AT2 cells (MLE12) treated for 24 h with conditioned medium from control macrophages ( n = 11) or those polarized to a pro-inflammatory ( n = 10) or anti-inflammatory ( n = 5) phenotype, normalized to β-actin and shown relative to control macrophages. q Western blot analysis of ALOX5 at 72 h in control macrophages ( n = 11) or those polarized to a pro-inflammatory ( n = 11) or anti-inflammatory ( n = 11) phenotype, normalized to β-actin and shown relative to control macrophages. r Western blot analysis of ACSL4 in AT2 cells (MLE12) treated for 24 h with conditioned medium from control macrophages or those polarized to a pro-inflammatory phenotype that were treated with siRNA against Alox5 , normalized to β-actin and shown relative to control macrophages, from individual experiments ( n = 3). Data shown mean ± SD, P values shown in charts determined by unpaired two-tailed Student’s t -test ( c , n ), two-tailed Mann–Whitney test ( e ), one-way ANOVA Bonferroni’s multiple comparisons test ( k , p , q ). Source data are provided as a Source Data file.

Article Snippet: The next day adherent cells were harvested, counted, and seeded at a density of 1 × 10 6 cells/ml in 24-well plates, and cultured for 24 h in fresh medium to obtain M0 cells, medium containing 1 μg/ml LPS (from Escherichia coli 0111:B4, Sigma-Aldrich) and 20 ng/ml recombinant murine IFNγ (ImmunoTools) for M1 or medium containing 20 ng/ml recombinant murine IL4 (ImmunoTools) for M2.

Techniques: Expressing, Western Blot, Immunohistochemical staining, Two Tailed Test, MANN-WHITNEY