Figure 4. Effects of casticin (CAS) on IL-1β–induced production of ICAM-1.

A549 cells (10⁶ cells/ml) were pretreated with CAS for 1 h and then stimulated with IL-1β (1 ng/ml) for 24 h to assay ICAM-1 protein expression by western blot (A), and ICAM-1 protein expressions were measured relative to the expression of β-actin (internal control) (B). ICAM-1 of the culture medium by ELISA (C). ICAM-1 gene expression levels, determined using real-time RT-PCR (D), and promoter assay were used to evaluate ICAM-1 promoter activity (E). THP-1 cells were labeled with calcein AM and co-cultured with A549 cells, followed by observation using fluorescence microscopy (F). Fluorescence intensity of THP-1 cell adhesion to A549 cells (G). The presented data are mean ± SEM; ^*p < 0.05, ^**p < 0.01, compared with the IL-1β–treated group.