anti-hil-1β-igg  (InvivoGen)

Journal: bioRxiv

Article Title: Dysplastic Epithelial Repair Propagates Chronic Pathology Through the Paracrine Transformation of Pulmonary Fibroblasts

doi: 10.64898/2026.04.02.716135

Figure Lengend Snippet: ( A ) Quantified levels of IL-1α from naïve fibroblast (Naïve fibro), inflammatory fibroblast (Inflam fibro), and ectopic basal cell (Basal) conditioned media as determined by ELISA. ( B ) Saa3 and Ccl2 gene expression by RT-qPCR for fibroblasts cultured on 0.5kPa plates and treated with a 5-fold titration of IL-1α added to control media, including the level determined in (A). ( C ) Graphic illustrating the strategy used to deplete conditioned media of endogenous IL-1α with antibodies, magnetic beads, and column-based separation. ( D ) Saa3 and Ccl2 gene expression by RT-qPCR for fibroblasts cultured on 0.5kPa plates in control or conditioned media depleted using IgG (CTRL IgG or CM IgG) or IL-1α nAb (CTRL nAB, CM nAb) and supplemented with additional IL-1α nAb after depletion (+Spike). [(B) and (D)] Data shows mean fold change ± SD and are pooled from 3 independent experiments (n=3 mice). P values were calculated using either (B) a ratio paired t -test or (D) a one-way ANOVA with post-hoc Turkey multiple comparison test: * P < 0.05, ** P < 0.01, **** P < 0.0001, ns: not significant.

Article Snippet: The working concentration for the following reagents can be found in the text and figure legends: Human IL-1RA Recombinant Protein (IL1Ra) (PeproTech®, 200-01RA), Mouse IL-1 alpha Recombinant Protein, (PeproTech®, 211-11A), Mouse IL-1α Neutralizing Antibody (Invivogen, Anti-mIL-1α-mIgG1 clone 6H7, mIL-1α-mab9-02), Recombinant Mouse IgG1 Isotype Control Antibody (Invivogen, Anti-β-Gal-mIgG1 clone T9C6, bgal-mab9-02), Human TGF-β 1 Recombinant Protein (PeproTech®, 100-21), Human GDF15 Recombinant Protein (PeproTech®, 120-28C), Human VEGF Recombinant Protein (PeproTech®, 450-32), Human OPN Recombinant Protein (PeproTech®, 120-35), Murine CXCL1 Recombinant Protein (PeproTech®, 250-11), Human IGFBP3 Recombinant Protein (PeproTech®, 100-08), Human TNFRSF11B Recombinant Protein (PeproTech®, 450-14), Murine CXCL5 Recombinant Protein (PeproTech®, 250-17), and Muring CCL20 Recombinant Protein (PeproTech®, 250-27).

Techniques: Enzyme-linked Immunosorbent Assay, Gene Expression, Quantitative RT-PCR, Cell Culture, Titration, Control, Magnetic Beads, Comparison

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