human ifnɣ  (R&D Systems)

Journal: Mobile DNA

Article Title: An intronic LINE-1 regulates IFNAR1 expression in human immune cells.

doi: 10.1186/s13100-023-00308-3

Figure Lengend Snippet: Fig. 4 IFNAR1.L1M2a inducibly enhances transcription of IFNAR1 during type I IFN signaling. A IFNAR1 and three nearby genes, IFNAR2, IL10RB, and IFNGR2, were all considered to be possible targets of IFNAR1.L1M2a enhancer activity due to their proximity. B RNA sequencing of three wildtype clones (gray) and four IFNAR1.L1M2a.enh knockout clones (purple) for IFNAR1.L1M2a-proximal genes during a 24 h time course of IFNβ treatment. Pairwise differential expression analysis [48] at the untreated timepoint showed slightly but significantly lower baseline expression of IFNAR1 and IFNGR2 in knockout cells, and likelihood ratio tests [48] showed significant differences in response to IFNβ treatment of IFNAR1 and IL10RB. At the 24 h timepoint, IFNAR1 expression was slightly but significantly upregulated compared to the untreated timepoint in wildtype cells but not in knockout cells, and expression of IFNAR1 was significantly lower in knockout cells. C-D Immunofluorescence was used to confirm protein-level differences in IFNAR1 expression between wildtype cells (left, gray) and knockout cells (right, purple) in untreated (top, light) and IFNβ-treated (bottom, dark) conditions. In agreement with RNA-seq data, IFNAR1 expression was induced by IFNβ treatment in wildtype cells, but not in knockout cells. Scale bar indicates 25 microns

Article Snippet: Cells were either untreated or were cultured in media containing 1000 U/ml human IFNɣ (#485-MI, R&D Systems) for 4 h before RNA extraction.

Techniques: Activity Assay, RNA Sequencing, Clone Assay, Knock-Out, Quantitative Proteomics, Expressing, Immunofluorescence

Journal: Mobile DNA

Article Title: An intronic LINE-1 regulates IFNAR1 expression in human immune cells.

doi: 10.1186/s13100-023-00308-3

Figure Lengend Snippet: Fig. 5 Deletion of IFNAR1.L1M2a.enh alters downstream transcriptional response to IFNβ. RNA sequencing of three wildtype (gray) and four IFNAR1.L1M2a.enh knockout clones (purple) during a 24 h time course of IFNβ treatment. A Likelihood Ratio Tests [48] of RNAsequencing data revealed many genes with significantly altered IFNβ responses in knockout cells compared to wildtype. Many of these genes were expected IFN stimulated genes (red) as identified by induction in wildtype cells at the 4 h timepoint [48] (Additional file 7: Table S6). Gene ontology [51] confirmed that significantly differentially responsive genes were enriched for immune-related biological processes (red). B RNA sequencing of representative expected ISGs showed a dampened initial induction in the knockout cells (purple) compared to wildtype cells (gray) and differential expression throughout the time course of IFNβ treatment according to likelihood ratio tests [48]

Article Snippet: Cells were either untreated or were cultured in media containing 1000 U/ml human IFNɣ (#485-MI, R&D Systems) for 4 h before RNA extraction.

Techniques: RNA Sequencing, Knock-Out, Clone Assay, Quantitative Proteomics

Journal: Cell Genomics

Article Title: Proteomics of immune cells from liver tumors reveals immunotherapy targets

doi: 10.1016/j.xgen.2023.100331

Figure Lengend Snippet:

Article Snippet: Recombinant human IFNɣ , Sino Biological , Cat#11725-HNAS.

Techniques: Recombinant, Sequencing, Modification, Transfection, DNA Extraction, Enzyme-linked Immunosorbent Assay, Cell Isolation, Expressing, Mouse Assay, Software