Facp array software v3

We measured the pro-inflammatory cytokines released from M1-macrophage-like THP-1 cells 48 h after C. concisus infection in co-culture cell monolayers. The media from the basal compartment of the C. concisus-infected and control cell monolayers were collected from 12 well plates. This was subjected to analysis of multiple cytokines (IL-1β, IL-13, IL-4, IL-6, IL-10, IL-17A, IFN-γ, and TNF-α) with capture bead technology of human cytometric bead array kit (CBA; BD Biosciences Human Th1/ Th2/Th17 Kit and Flex Set IL-13 and IL-1β, Franklin Lakes, NJ, USA) according to the manufacturer’s protocol. The cytokines were then measured with flow cytometry by Fluorescence-activated Cell Sorting (FACS) Canto II (BD Biosciences; Franklin Lakes, NJ, USA). The data from the cytokine measurements were analyzed with FACP Array software v3.0 (BD Biosciences, Franklin Lakes, NJ, USA).

At the timepoint of 22 h after infection we collected the medium from basal compartment of the 12 well plate to analyze the secretion of cytokines (IL-1β, IL-2, IL-4, IL-6, IL-10, IL-17A, IFN-γ, and TNF-α) during the infection using a human cytometric bead array kit and manufacturers protocol (CBA; BD Biosciences human Th1, Th2, Th17 Kit, Flex Set IL-1β, Franklin Lakes, NJ, United States). Flow cytometric measurement were performed with FACS CantoII (BD Biosciences; Franklin Lakes, NJ, United States) and analyzed with FACP Array software v3.0 (BD Biosciences, Franklin Lakes, NJ, United States).