Fig. 1: Effects of IL-38 on the cytokines/chemokines released in co-cultured human respiratory epithelial cells with HMDMs.
A549 cells (1 × 105) and/or BES-2B cells (1 × 105) with HMDMs (3 × 105) were co-cultured with or without rhIL-38 (100 ng/ml) pretreatment for 30 min, followed by poly(I:C) (20 µg/ml) stimulation for 20 h. (a–k) Release of cytokines/chemokines in the co-cultured supernatants were measured by Cytometric Bead Array Flex Sets using flow cytometry. (l) Gating strategy of flow cytometry for the determination of ICAM-1. (m) Flow cytometric analysis of cell surface expression of ICAM-1 on the monoculture of A549 cells, BEAS-2B cells and HMDMs. (n, o) ICAM expression on A549 cells (Co-A549) in the co-cultured A549 cells with HMDMs, on BEAS-2B cells (Co-BEAS-2B) in the co-cultured BEAS-2B cells with HMDMs, and on HMDMs (Co-HMDMs) in the co-cultured A549/BEAS-2B cells with HMDMs. A549 cells, BEAS-2B cells, and HMDMs in the co-cultures were gated based on CD45 expression. Heat-inactivated (56 °C for 30 min.) human IL-38 (100 ng/ml) was used as the control. Abbreviations: HMDMs, human monocytes derived macrophages; Co-A549, A549 cells in the co-cultured A549 cells with HMDMs; Co-BEAS-2B, BEAS-2B cells in the co-cultured BEAS-2B cells with HMDMs; Co-HMDMs, HMDMs in the co-cultured A549/BEAS-2B cells with HMDMs. All data are shown as the mean ± SEM. Nonparametric Kruskal-Wallis test followed by Dunn’s multiple comparisons test was used to compare results between groups. *P < 0.05 and **P < 0.01.
