ligand fsl 1  (InvivoGen)

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: Cells were then washed, trypsinized, and seeded onto the labeled macrophages along with the macrophage-activating ligand FSL-1 (1 ng/ml) (Invivogen, #tlrl-fsl) overnight.

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

Journal: Materials Today Bio

Article Title: Type 1 diabetes mellitus (T1DM) does not affect whole blood responses to alginate-based microspheres despite plasma lipid and glucose differences

doi: 10.1016/j.mtbio.2025.102113

Figure Lengend Snippet: Immune functional testing using distinct TLR ligands (TLR2, TLR3, TLR4, TLR7, TLR8) ligands for Dectin and STING, and a T cell stimulus with human whole blood from T1DM and healthy controls. Multiple immune response parameters (n = 11) in blood from T1DM patients (n = 19) and controls (n = 19) after ex-vivo treatment with distinct TLR ligands were analysed by Principal Component Analysis (PCA) to compare the study groups. (A) Response to TLR ligands visualised as a PC1 vs PC2 plot. (B) Dot plot and statistical testing of individual PC (PC1 to PC5) scores, showing the overall ANOVA and pairwise post-test p-values, which do not reveal any significant differences between the groups. The variance of the original data that the PCs cover is shown below. (C) Volcano plots comparing the induction of response variables in the groups by distinct TLR ligands, revealing no significant differences.

Article Snippet: For the immune functional assay (IFA) blood model (section ) immune agonists from InvivoGen were used at the following final concentrations: TLR2 ligand (FSL-1, 100 ng/mL), TLR3 ligand (poly I:C, 20 μg/mL), TLR4 ligand (LPS O111:B4, 1 ng/mL), TLR7 ligand (Gardiquimod, 1 μg/mL), TLR8 ligand (TL8-506, 300 ng/mL), dectin ligand (whole glucan particle (WGP®, 40 μg/mL), and STING ligand (2′3′- cGAMP, 13 μg/mL).

Techniques: Functional Assay, Ex Vivo