Protocol for high-throughput compound screening using flow cytometry in THP-1 cells

• Published in: STAR protocols Vol. 2; no. 2; p. 100400
• Main Authors: Spangenberg, Stephan H., Zavareh, Reza Beheshti, Lairson, Luke L.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 18.06.2021; Elsevier
• Subjects: Cell-based assays; Flow cytometry/mass cytometry; High-throughput screening; Immunology; Protocol; High-throughput screening; Immunology; Cell-based assays; Flow cytometry/mass cytometry
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2021.100400

Flow cytometry is a valuable method for analyzing protein expressions at the single cell level but can be difficult to apply to large numbers of samples. This protocol provides instructions to perform a high-throughput small molecule screen using flow cytometry analysis of THP-1 cells, a human monocytic leukemia cell line. We describe a methodology for identifying compounds that regulate PD-L1 surface expression in IFN-γ-stimulated cells, which has been successfully used to screen a collection of ∼200,000 compounds. For complete details on the use and execution of this protocol, please refer to Zavareh et al. (2020). [Display omitted] •Protocol for high-throughput screening of compounds using flow cytometry•Designed for quantification of cell surface expression of proteins in THP-1 cells•This protocol has been used to identify modulators of PD-L1 expression Flow cytometry is a valuable method for analyzing protein expressions at the single cell level but can be difficult to apply to large numbers of samples. This protocol provides instructions to perform a high-throughput small molecule screen using flow cytometry analysis of THP-1 cells, a human monocytic leukemia cell line. We describe a methodology for identifying compounds that regulate PD-L1 surface expression in IFN-γ-stimulated cells, which has been successfully used to screen a collection of ∼200,000 compounds.