Ifn gamma

Manufactured by Cell Signaling Technology

IFN-gamma is a recombinant cytokine that functions as an interferon. It is involved in the regulation of immune responses and plays a role in various cellular processes.

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Lab products found in correlation

Picopipet, by Nepa Gene (1 mentions) Evos fluorescent microscope, by Thermo Fisher Scientific (1 mentions) Nunc lab tek 2 chamber slide system, by Thermo Fisher Scientific (1 mentions)

2 protocols using ifn gamma

Establishment and Characterization of Erlotinib-Resistant Lung Cancer Cells

Cited 1 time

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Human lung cancer cell lines used in this study were from the established collections in our labs or as reported in our previous studies [8 (link)–10 (link)]. PC-9 erlotinib resistant cells were established from PC-9 cells by stepwise exposure to erlotinib from 0.005 μM to 5 μM for about 4 months, and the clone named PC-9 ER clone 5 was isolated with PicoPipet (Nepa Gene, Chiba, Japan). Cells were cultured in RPMI1640 medium supplemented with 10% fetal bovine serum (FBS) and 1x penicillin/streptomycin solution (Mediatech, Inc., Manassas, VA) at 37°C in a humidified tissue culture incubator with 5% CO₂. All experiments using acquired resistance cells were performed following the removal of drug exposure to avoid the direct effects of drugs on PD-L1 expression. IFN-gamma (Cell Signaling Technology, Dancers, MA) stimulation for 24 hours was performed to mimic an immune cell interaction.

Suda K., Rozeboom L., Furugaki K., Yu H., Melnick M.A., Ellison K., Rivard C.J., Politi K., Mitsudomi T, & Hirsch F.R. (2017). Increased EGFR Phosphorylation Correlates with Higher Programmed Death Ligand-1 Expression: Analysis of TKI-Resistant Lung Cancer Cell Lines. BioMed Research International, 2017, 7694202.

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Immunofluorescence Assay for E-cadherin and PD-L1

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Cells were seeded in an 8-well Nunc Lab-Tek II Chamber Slide System (Thermo Scientific, Rochester, NY), and treated the day after with PBS or IFN-gamma (Cell Signaling Technology) for 24 hours. Immunofluorescence was performed according to the manufacture’s protocol. Briefly, cells were fixed with 4% formaldehyde, blocked with blocking buffer, and incubated with primary antibodies for E-cadherin (mouse monoclonal) or PD-L1 (rabbit monoclonal) at 4°C overnight. After the washing with PBS, cells were incubated with fluorochrome-conjugated secondary antibodies for anti-rabbit and anti-mouse (Cell Signaling Technology). Cell nuclei were stained with 4’,6-diamidino-2-phenylindole (DAPI). Microscopy was performed on an EVOS fluorescent microscope (Model FL, Life Technologies, Carlsbad, CA).

Suda K., Rozeboom L., Rivard C.J., Yu H., Ellison K., Melnick M.A., Hinz T.K., Chan D., Heasley L.E., Politi K., Mitsudomi T, & Hirsch F.R. (2017). Therapy-induced E-cadherin downregulation alters expression of programmed death ligand-1 in lung cancer cells. Lung cancer (Amsterdam, Netherlands), 109, 1-8.

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