miR-29s suppress cytokine-induced expression of sST2 and IFNAR1 in BEAS-2B cells
A, Cytokine-induced soluble ST2 (sST2) release from BEAS-2B cells. A human bronchial epithelial cell line, BEAS-2B cells, was stimulated with the indicated concentrations of IL-4 and TNF-α, alone or in combination, for 48 h. The protein concentration of sST2 in each culture supernatant was measured by ELISA. B, Schematic representation of the gene structures of miR-29 family members. C, The effects of inhibition or overexpression of miR-29s on sST2 release from BEAS-2B cells were examined. Following 24-h transfection of 10 nM of miRNA for miR-29 inhibitors, mimics or a negative control, the cells were stimulated with a combination of 100 ng/ml IL-4 and TNF-α for 48 h. The protein concentration of sST2 in each culture supernatant was measured by ELISA, and the data are shown as the mean ± SD of triplicate samples. *P< .05 by Student’s t -test. D, Effects of inhibition or overexpression of miR-29s on the expression of ST2 and IFNAR1 proteins in BEAS-2B cells. BEAS-2B cells were treated as above in C. Whole-cell lysates were subjected to Western blot analysis of the expression of ST2, IFNAR1 and β-actin (as a loading control). The data are representative of two separate experiments. Chr.: chromosome; NC: negative control; IFNAR1: IFN (α, β and ω) receptor 1; ACTB: β-actin.