recombinant human interferon α 2a  (MedChemExpress)

Journal: EMBO Reports

Article Title: Type I interferon exacerbates Mycobacterium tuberculosis induced human macrophage death

doi: 10.1038/s44319-024-00171-0

Figure Lengend Snippet: ( A ) IFN-β or ( B ) IFN-α were added to Mtb H37Rv infected MDM-1 ( n = 3) at 4 h post infection after extracellular Mtb was washed out. Cell viability was determined by CellTiter Glo assay at 2 d post infection. MDM-1 were primed with ( C ) IFN-β or ( D ) IFN-α; 1 day later MDM-1 ( n = 3) were infected with Mtb H37Rv. Cell viability was measured by CellTiter Glo assay at 2 d post infection. Cell viability was normalized to values for the respective uninfected, unstimulated or non-primed MDMs. The fold change of the normalized cell viability was calculated by dividing the normalized cell viability of IFN-I stimulated MDMs by that of unstimulated MDMs within the same group (uninfected or infected) per donor. ( E ) Images of MDM-1 ( n = 3) primed with IFN-β 1 day pre-infection, infected with Mtb H37Rv, and imaged at 20x at 2 d post infection. Scale bars = 20 μm. Data Information: ( A – D ) Bar graphs report the mean ± SEM of the fold change. Each ● represents the average value of 3 technical replicates per donor ( n = 3), n indicates biological replicates. Cells from each donor were tested in independent experiments. Statistical significance was determined using a paired, two-tailed t -test (* p < 0.5; ** p < 0.01; n.s. indicates no significance). .

Article Snippet: Recombinant human IFN-α 2a , PBL Assay Science , Cat # 11100-1.

Techniques: Infection, Glo Assay, Two Tailed Test

Journal: EMBO Reports

Article Title: Type I interferon exacerbates Mycobacterium tuberculosis induced human macrophage death

doi: 10.1038/s44319-024-00171-0

Figure Lengend Snippet: ( A ) Representative images of MDM-1 ( n = 1) that were primed with IFN-α 1 day pre-infection, infected with Mtb H37Rv, stained with 7-aminoactinomycin D (7-AAD) and imaged at 10×2 d post infection. Scale bars = 100 μm. ( B ) MDM-2 ( n = 1) were primed with IFN-β 1 day pre-infection, infected with Mtb H37Rv, and cell viability was measured via CellTiter Glo assay. Data Information: ( B ) Dot plot reports mean value of 3 technical replicates per 1 donor, n indicates biological replicates.

Article Snippet: Recombinant human IFN-α 2a , PBL Assay Science , Cat # 11100-1.

Techniques: Infection, Staining, Glo Assay

Journal: EMBO Reports

Article Title: Type I interferon exacerbates Mycobacterium tuberculosis induced human macrophage death

doi: 10.1038/s44319-024-00171-0

Figure Lengend Snippet: Reagents and tools table

Article Snippet: Recombinant human IFN-α 2a , PBL Assay Science , Cat # 11100-1.

Techniques: Recombinant, Sequencing, Modification, Saline, Enzyme-linked Immunosorbent Assay, Cell Viability Assay, Membrane, Sterility, Pore Size, Western Blot, Stripping, Software, Spectrophotometry, Microscopy, Microplate Reader Luminescence Measurement, Autoradiography, Imaging

Journal: Arthritis Research & Therapy

Article Title: The potential roles of type I interferon activated neutrophils and neutrophil extracellular traps (NETs) in the pathogenesis of primary Sjögren’s syndrome

doi: 10.1186/s13075-022-02860-4

Figure Lengend Snippet: Results for rhIFN-α stimulation of neutrophils in pSS patients and healthy controls. A Results MitoSOX measurements of pSS and HC plasma stimulation of healthy neutrophils ( n =9). B Representative flow cytometry results for the JC-1 staining in pSS patients and healthy controls with or without rhIFN stimulation (HC\pSS-Unstimulated represented for the freshly isolated neutrophils, pSS\HC-with or without rhIFN represented for the neutrophils stimulated with or without rhIFN). C Representative flow cytometry results of ROS production in pSS and healthy neutrophils with or without rhIFN stimulation (HC\pSS-Unstimulated represented for the freshly isolated neutrophils, pSS\HC-with or without rhIFN represented for the neutrophils stimulated with or without rhIFN, different color represented for the different subjects). D Comparison of JC-1 monomer percentage (mitochondrial damage, n =10), ROS production ( n =10), and MPO concentration ( n =12) between neutrophils stimulated with or without rhIFN-α. E Immunofluorescence staining of NETs-DNA (DAPI, blue) with or without rhIFN stimulation. F IFN-α receptor inhibitor reduced the mitochondrial damage (MitoSOX) and the production of NETs (SytocGreen and MPO levles). * P -value < 0.05, ** P -value < 0.01, *** P -value <0.001)

Article Snippet: As for the plasma stimulation experiment, we firstly pooled the plasma from 15 treatment-naïve pSS patients and 15 matched HCs together, then prepared RPMI 1640 medium (Gibco, USA) combined with 20% pSS or HC plasma, and then cultured with the freshly isolated neutrophils for 4 h. To further confirm the effect of rhIFN-α 2a on neutrophils, freshly isolated neutrophils were pretreated with IFN-α receptor inhibitor [IFN alpha-IFNAR-IN-1 hydrochloride, MedChemexpress (MCE), USA] for half an hour and then stimulated with rhIFN-α 2a for another 4 h.

Techniques: Clinical Proteomics, Flow Cytometry, Staining, Isolation, Comparison, Concentration Assay, Immunofluorescence