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. 2023 Jan 8;24(2):1227. doi: 10.3390/ijms24021227

Figure 5.

Figure 5

Montelukast upregulated IL25, IL33, and TSLP expression via histone acetylation. A549 cells were pretreated with anacardic acid (AA) for 1 h and stimulated with montelukast for another 3 h. The supernatant for IL-25, IL-33, and TSLP protein measurement were collected after treatment for 24 h in A549 cells or 96 h in ALI cultures. The montelukast-induced changes in the mRNA (A–C), (n = 5) or protein (D–F), (n = 4) expression of IL-25, IL-33, and TSLP were suppressed by AA. For ChIP assays, the cells were treated with montelukast for 0.5 h before the DNA was extracted. Montelukast increased the acetylation of histone H3, but not of histone H4, at the IL25 (G), IL33 (H), and TSLP (I) promoter regions (n = 4). The montelukast-induced changes in the protein expression of IL-25, IL-33, and TSLP were suppressed by AA in ALI cultures (J), (n = 4). Montelukast increased the acetylation of histone H3 at the IL25 (K), IL33 (L), and TSLP (M) promoter regions (n = 3). IL25, IL33, and TSLP mRNA expression, histone acetylation in the promoter region, or protein levels were detected using real-time PCR or ELISA. means ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001 compare with solvent control (Con); # p < 0.05, ## p < 0.01, and ### p < 0.001 compare with montelukast alone (Mon).