Thp 1

BEAS-2B cells were treated with various concentrations of sesamol and stimulated with or without TNF-α/IL-4 for 24 h. Human monocytic cells (THP-1, Bioresource Collection and Research Center, Taiwan) were cultured in RPMI 1640 medium. THP-1 cells were stained with calcein-AM solution (Sigma). Next, THP-1 cells were co-cultured with BEAS-2B cells, and adherent THP-1 cells were evaluated with fluorescence microscopy (Olympus, Tokyo, Japan), as previously described [21 (link)].

For activation, HMEC-1 cells (human microvascular endothelial cell-1) were cultured in 24-well plates and incubated with LPS and metformin in the presence or absence of NAM for 24 h. For monocyte–ECs adhesion experiments, assays were performed by adding 5  ×  10⁵ THP1 cells (Zhong Qiao Xin Zhou Biotechnology Co., Ltd., Shanghai, China) that were labeled with 2 µM calcein-AM with green fluorescence (Beyotime Biotechnology, China) to 1  ×  10⁵ confluent HMEC-1 cells (FuHeng Biology, Shanghai, China) for 2  h (Lin et al., 2018 (link)). THP-1 cells that were firmly bound to HMEC-1 were observed with a fluorescence microscope (Olympus, Tokyo, Japan).