il-28r alpha/ifn-lambda r1 antibody  (Bio-Techne corporation)

Journal: Frontiers in Immunology

Article Title: Intracellular Accumulation of IFN-λ4 Induces ER Stress and Results in Anti-Cirrhotic but Pro-HCV Effects

doi: 10.3389/fimmu.2021.692263

Figure Lengend Snippet: IFN-λ4 inhibits the proliferation of HepG2 cells. RNA-sequencing analysis was used to identify differentially expressed genes (DEGs, P-FDR < 0.05) in IFN-λ3-GFP, IFN-λ4-GFP and IFN-λ4-GFP-IFNLR1 KO HepG2 cells after 72 hrs of induction by dox, compared to controls (dox- conditions). The cutoff threshold (fold change > +/-1.5) is indicated by dotted lines. (A) Analysis of all DEGs (n = 3251) detected for IFN-λ4-GFP or IFN-λ4-GFP-IFNLR1 KO cells. In blue - DEGs (n = 2,735) specific to IFN-λ4-GFP and considered IFNLR1-dependent. In black - DEGs (n = 145) shared between both groups and considered IFNLR1-independent. In orange - DEGs (n = 371) specific to IFN-λ4-GFP-IFNLR1 KO . (B) DEGs of IFN-λ4-GFP analyzed in IFN-λ3-GFP transcriptome. In black - DEGs (n = 1,506) shared in IFN-λ4-GFP and IFN-λ3-GFP and in blue - IFN-λ4-signature DEGs (n = 1,229) detected in IFN-λ4-GFP but not in IFN-λ3-GFP producing cells. Additional details are provided in and . (C, D) Cell cycle analysis of cells synchronized by 24 hrs of serum starvation, treated with or without dox (0.5 µg/ml) for 72 hrs, and analyzed by flow cytometry after PI staining. The plot shows a representative picture and the percentage of cells in each phase of the cell cycle. All data are shown as mean± SEM from triplicate experiments. *P < 0.05, Student’s T-test comparing no dox to dox+ for each cell cycle stage. (E, F) Bromodeoxyuridine (BrdU, %) incorporation indicating cell proliferation in HepG2 cells expressing IFN-λ3-GFP, IFN-λ4-GFP and IFN-λ4-GFP-IFNLR1KO. Cells were cocultured with HepG2 cells labeled with Far Red proliferation dye, dox-induced for 72 hrs, and treated with BrdU for 3 hrs before analysis. Gates show HepG2 cells exposed to IFN-λs (IFN-λ treated cells) and HepG2 expressing IFN-λs. P-values compare corresponding dox+ vs. dox- HepG2 cells, **p < 0.01, Student’s T-test. Graphs represent one of three independent experiments, each in biological triplicates.

Article Snippet: The membranes were probed with primary antibodies against IFNLR1 (#NBP1-84381, Novus Biologicals), STAT1 (#9172, Cell Signaling Technology), phospho-STAT1 (Tyr701, #58D6, Cell Signaling Technology), IFN-λ4 (ab196984; Abcam), GAPDH (ab37168, Abcam) and HRP-linked secondary antibody, goat anti-rabbit IgG (#7074; Cell Signaling Technology) or goat anti-mouse IgG (San Cruz, sc-2031).

Techniques: RNA Sequencing, Cell Cycle Assay, Flow Cytometry, Staining, BrdU Incorporation Assay, Expressing, Labeling

Journal: Frontiers in Immunology

Article Title: Intracellular Accumulation of IFN-λ4 Induces ER Stress and Results in Anti-Cirrhotic but Pro-HCV Effects

doi: 10.3389/fimmu.2021.692263

Figure Lengend Snippet: IFN-λ4 is a misfolded protein that induces ER stress. Representative confocal images of HepG2 cells transduced with a mammalian baculovirus delivery system (BacMam) of GFP-tagged proteins targeting specific organelles - lysosomes, Golgi, early and late endosomes. After transduction for 6 hrs, cells were transiently transfected with Halo-tagged constructs for IFN-λ4 or control for indicated times, stained with cell-permeant Halo-tag ligand TMR (red), and imaged. (A) Confocal images showing IFN-λ4 accumulation in lysosomes but not in early endosomes. (B) Late endosomal trafficking of IFN-λ4, with the inset showing larger magnification. (C) Unfolded protein response (UPR) is represented by lysosomal enlargement after protein accumulation. (D) Live images of IFN-λ4-expressing HepG2 cells undergoing apoptosis, characterized by membrane blebbing and cell death. Images were scanned every minute for 12 hrs. Scale bars – 10 um. (E) Apoptosis detection with ApoTox-Glo assays in corresponding untreated and dox-induced cells for indicated time points. RLU, relative luminescence units. (F) Graph showing counts from colony formation assay for HepG2 cells expressing IFN-λ3, IFN-λ4 or IFNLR1 KO grown in 6-well plates with or without dox for 13 days. Cell colonies were stained with crystal violet and counted with ImageJ software. The graph represents the number of colonies as a percentage of initial plated counts. n.s., not significant, ***p < 0.001, ****p < 0.0001, Student’s T-test.

Article Snippet: The membranes were probed with primary antibodies against IFNLR1 (#NBP1-84381, Novus Biologicals), STAT1 (#9172, Cell Signaling Technology), phospho-STAT1 (Tyr701, #58D6, Cell Signaling Technology), IFN-λ4 (ab196984; Abcam), GAPDH (ab37168, Abcam) and HRP-linked secondary antibody, goat anti-rabbit IgG (#7074; Cell Signaling Technology) or goat anti-mouse IgG (San Cruz, sc-2031).

Techniques: Transduction, Transfection, Construct, Control, Staining, Expressing, Membrane, Colony Assay, Software

Journal: Clinical and Experimental Immunology

Article Title: Expression of type III interferons (IFNλs) and their receptor in Sjögren's syndrome

doi: 10.1111/cei.12865

Figure Lengend Snippet: Expression of type III interferons (IFNs) (IFN‐λ1/interleukin (IL)−29, IFN‐λ2/IL‐28A and IFN‐λ3/IL‐28B) and their signalling receptor (IFN‐λR1/IL‐28Rα) (a) in minor salivary gland (MSG) tissues that belong to the three Sjögren's syndrome (SS) subgroups, as these classified by the grade of the inflammatory lesion (SS‐I: mild, n = 16; SS‐II: intermediate, n = 14; and SS‐III: severe, n = 16 MSG lesions. Sicca controls CT, n = 17), (b) in non‐malignant parotid glands of patients undergoing parotidectomy due to mixed tumours. Original magnification ×200.

Article Snippet: Rabbit polyclonal antibodies to human IFN‐λ1/IL‐29 (Santa Cruz, Dallas, Texas, USA), IFN‐λ2/IL‐28A (Antibodies‐online, Atlanta, GA, USA), IFN‐λ/IL‐28B (Bioss, Woburn, MA, USA), IFN‐λR1/IL‐28Ra (Novus Biologicals, Littleton, CO, USA) and CD3 (Dako, Glostrup, Denmark) were used.

Techniques: Expressing

Journal: Clinical and Experimental Immunology

Article Title: Expression of type III interferons (IFNλs) and their receptor in Sjögren's syndrome

doi: 10.1111/cei.12865

Figure Lengend Snippet: Representative double immunofluorescent staining for the common type III interferon (IFN) signalling receptor [IFN‐λR1/interleukin (IL)−28Rα] and markers of inflammatory cells [plasmatocytoid dendritic cells (pDCs): CD303 and CD123, macrophages: CD68, T lymphocytes: CD3 and B lymphocytes: CD20] in a minor salivary gland (MSG) tissue from a Sjögren's syndrome (SS) patient. Co‐localization is indicated with yellow arrows. Original magnification ×1000.

Article Snippet: Rabbit polyclonal antibodies to human IFN‐λ1/IL‐29 (Santa Cruz, Dallas, Texas, USA), IFN‐λ2/IL‐28A (Antibodies‐online, Atlanta, GA, USA), IFN‐λ/IL‐28B (Bioss, Woburn, MA, USA), IFN‐λR1/IL‐28Ra (Novus Biologicals, Littleton, CO, USA) and CD3 (Dako, Glostrup, Denmark) were used.

Techniques: Staining