interferon α 2  (MedChemExpress)

Journal: Molecular Oncology

Article Title: Enhancing immunotherapy through PD ‐ L 1 upregulation: the promising combination of anti‐ PD ‐ L 1 plus m TOR inhibitors

doi: 10.1002/1878-0261.13699

Figure Lengend Snippet: Possible mechanisms by which mTOR inhibitors might regulate PD‐L1. (A, B) Cells were seeded in 100 mm 2 and 24 h later, cells were treated with everolimus (100 n m ) or TAK‐228 (50 n m ) for 48 h. (A) Effect of the drugs on S6 and GSK3 protein expression. Cellular extracts were analyzed by western blot using the indicated antibodies. The quantifications show the ratio between phosphorylated and total protein in each condition, expressed as fold induction versus control arbitrarily set at 1 from three independent experiments. (B) Effects of the drugs on the mRNA expression of EGF or IFNβ in T24 (top) and CAL‐29 (below). The mRNA levels of EGF or IFNβ were analyzed by qRT‐PCR after the treatment with the drugs. mRNA levels were normalized to the control cells expression level set at 1. The average of three independent experiments is shown. (C, D) Effects of the blockage of EGFR by cetuximab in CAL‐29 cells. Cells were treated with cetuximab (15 μg·mL −1 ) for 3 h and then TAK‐228 (50 n m ) or EGF (25 ng·mL −1 ) were added for 48 h. (C) Cellular extracts were analyzed by western blot. The graph shows the levels of PD‐L1 expressed as fold induction versus control arbitrarily set at 1 from three independent experiments. The black dot indicates the glycosylated form of PD‐L1. (D) The graph shows the fold‐change MFI ratio (mean fluorescence intensity ratio) for PD‐L1 relative to control cells. Representative image of the PD‐L1 expression detected by flow cytometry from three independent experiments. (E) Effects of the blockage of IFNAR by anti‐IFNAR2 in CAL‐29 cells. Cells were treated with anti‐IFNAR2 (0.4 μg·mL −1 ) for 3 h and then TAK‐228 (50 n m ) and/or IFNβ (2700 U) was added for 48 h. Cellular extracts were analyzed by western blot. Values show the levels of PD‐L1 expressed as fold induction versus control arbitrarily set at 1 from two independent experiments. The black dot indicates the band of the glycosylated form of PD‐L1. (F) Effects of the dual blockage of EGFR and IFNAR in CAL‐29 cells. Cells were treated with anti‐IFNAR2 (0.4 μg·mL −1 ) and cetuximab (15 μg·mL −1 ) for 3 h and then TAK‐228 (50 n m ) was added for 48 h. Cellular extracts were analyzed by western blot. Values show the levels of PD‐L1 expressed as fold induction versus control arbitrarily set at 1 from three independent experiments. The black dot indicates the band of the glycosylated form of PD‐L1. (G) Schematic representation of the proposed mechanism of action for TAK‐228. Illustration was created with BioRender.com . Error bands indicate standard deviation (SD). Asterisks indicate significant differences between groups by Student t ‐test (* P < 0.05, ** P < 0.01 and *** P < 0.001).

Article Snippet: Anti‐αIFNAR2: Anti‐α‐Interferon Receptor Chain 2 Mouse mAB (MMHAR‐2) (407295; Merck‐Life Science, Darmstadt, Germany) was dissolved at 500 μg·mL −1 in PBS 0.1% BSA and was stored at −80 °C.

Techniques: Expressing, Western Blot, Control, Quantitative RT-PCR, Fluorescence, Flow Cytometry, Standard Deviation

Journal: PLOS ONE

Article Title: Development of rhesus macaque astrocyte cell lines supporting infection with a panel of viruses

doi: 10.1371/journal.pone.0303059

Figure Lengend Snippet: Cells seeded in 12-well plates were either treated with 100 U/mL pan-IFN or infected with VSV ncp* (MOI 0.1). Untreated cells served as control. Cells were harvested after 24 h for RNA isolation. Expression of (A) interferon beta (IFNB1) or (B) MX1 was analyzed by quantitative RT-PCR. Transcript levels were normalized against 18S rRNA transcript levels and expression fold change was calculated with respect to control cells. The results of a representative experiment carried out with technical triplicates are shown. Error bars indicate standard deviation. Similar results were obtained in a separate experiment. Statistical significance was tested by two-way ANOVA: *, p≤0.05; **, p≤0.01; ***, p≤0.001.

Article Snippet: For IFN treatment, universal type I IFN-α (pan-Interferon; 11200–2, PBL Assay Science, Piscataway), a chimeric interferon constructed from human IFN alpha A and alpha D [ ] was added at 100 U/mL.

Techniques: Infection, Control, Isolation, Expressing, Quantitative RT-PCR, Standard Deviation