Journal: Nature

Article Title: Bidirectional CRISPR screens decode a GLIS3-dependent fibrotic cell circuit

doi: 10.1038/s41586-025-09907-x

Figure Lengend Snippet: (a) Secreted IL-11 after co-culture of fibroblast knockouts for TGFB1 - and IL1B -related ligands or receptors with TLR2/6-activated macrophages (top) or knockouts in activated macrophage knockouts with fibroblasts (bottom). n=3 cell lines per condition. (b) Z-score heatmap of IAF genes in TGFBR1/2 , IL1R1 CRISPRko fibroblasts co-cultured with TLR2/6-activated macrophages, normalized to HPRT. n=3 cell lines per condition. (c) qPCR of gene knockouts normalized to HPRT . Macrophages: IL-4/IL-13 (10 ng/mL); FSL-1 (10 ng/mL) + ATP (5 mM). Fibroblasts: media. n=2 cell lines per condition. (d) Secreted TGF-β and IL-1β after co-culture of primary human macrophages and fibroblasts (24 hours) ( Methods ). n=3 cell lines per condition. (e) Secreted IL-11 from primary colonic fibroblasts stimulated with TGF-β and/or IL-1β (10 ng/mL, 24 hours). Dashed lines: additive or synergistic response ( Methods ). One-way ANOVA with Tukey’s multiple-comparisons test. n=3 cell lines per condition. (f) Left: immunofluorescence of Il11 mNG colons after intraperitoneal injection with IgG control or dual anti-TGF-β and anti-IL-1β antibodies (100 μL in PBS, 100 μg/mouse). Right: IL-11 mNG cell percentage in all DAPI-imaged cells from two pooled independent experiments. Mice (co-housed, 13-20 weeks) were treated with chronic DSS (2.0%, 35 days). n=7 mice per condition. (g) qPCR quantification of Il11 from lysates from (f) normalized to Eef2 . Kruskal-Wallis test with Dunn’s multiple-comparison test. (h) Total colonic collagen percentage quantification from (f). (i) Quantification of colonic hydroxyproline normalized to total protein from lysates from (f). (j) Percent starting weight of mice from (f). Filled lines represent s.e.m. Linear mixed-effects analysis with Dunnett’s multiple comparison test. (k) Histopathological scoring ( Methods ) of H&E-stained tissues from (f). (l) Colon length measurements from (f). Unless otherwise stated, statistics are by a one-way ANOVA with Dunnet’s multiple comparison test on distinct biological replicates and error bars are the mean ± s.e.m. ns, not significant.

Article Snippet: Il11 mNG mice were administered DSS following the chronic DSS model. At the start of each cycle of DSS administration, mice were IP injected with monoclonal antibodies (mAbs) directed against IgG (BioXCell, #BP0083), anti-TGF-β (BioXCell, #BP0057), anti-IL-1β (Invivogen, #mil1b-mab9-1T), or the combination of anti-TGF-β and anti-IL-1β.

Techniques: Co-Culture Assay, Cell Culture, Immunofluorescence, Injection, Control, Comparison, Staining

Journal: Cell Reports

Article Title: Platelets Fuel the Inflammasome Activation of Innate Immune Cells

doi: 10.1016/j.celrep.2020.107615

Figure Lengend Snippet:

Article Snippet: hIL1B-Fwd: 5′-tgggcagactcaaattccagct-3′ , Metabion , N/A.

Techniques: Blocking Assay, Control, Purification, Recombinant, Activity Assay, Protease Inhibitor, Cell Culture, Modification, Saline, Clinical Proteomics, Transfection, Antibody Labeling, Isolation, Bicinchoninic Acid Protein Assay, LDH Cytotoxicity Assay, Mass Spectrometry, Knock-In, Software