Fig. 4

Cytokine and chemokine secretions by MIECs. 1 × 10⁶ MIECs were stimulated with 10, 30, 50 and 100 µg of EfSEVs and PBS (0 µg of EfSEVs) for 24 h. Also, 50 µg of EfSEVs were used to stimulate 1 × 10⁶ MIECs for 6, 12, 18 and 0 h as experimental control. IL-6, IL-17, MCP1, IL-1β and IL-18 secretions in cell culture supernatants were measured by protein microarray kit. The signals of the laser scanning map were extracted with GenePix 6.0 microarray analysis software. Quantitative data obtained from the Quantibody-INF-1Q-Analyzer were analyzed using RayBiotech mouse Inflammation Array 1 software. Cytokine concentrations (pg/ml) were determined by mean fluorescence intensities and linear regression standard curves were generated from the manufacturer’s standard. Each spot on the graph represents quantitative mean ± SD values in triplicate experiments. Asterisks indicate statistical significant differences from the control treatments at: *P < 0.05, **P < 0.001, ***P < 0.0001. Abbreviations: IL, Interleukin; MCPI, monocyte chemoattractant protein 1; MIECs, mouse intestinal epithelial cells; PBS, phosphate-buffered saline