Hts fluoroblok 96 multiwell insert system chemotaxis chambers

THP-1 cells stably expressing either GPSM3-knockdown shRNA or scrambled shRNA control were serum-starved in HBSS + 1% BSA at 37°C for one hour and labeled with calcein-AM cell permeant fluorescent dye (494 nm/517 nm). Cells were washed and suspended in HBSS + 1% BSA + 10 mM HEPES, and 5 × 10⁷ cells/ml were loaded into the upper well of a Falcon™ HTS FluoroBlok 96-Multiwell Insert System chemotaxis chambers (Corning, NY). Cells were allowed to migrate toward vehicle or 100 ng/mL MCP-1/CCL2 (R&D Systems, Minneapolis, MN) in the lower chamber. Fluorescence was measured at 2 minute intervals over the course of 60 minutes at 37°C using a Flexstation 3 microplate reader (Molecular Devices, Sunnyvale, CA). All data are representative of biological triplicates with technical replicates (i.e., n = 6).