Figure 2.

Glucocorticoids regulate the cell migration of macrophage-like THP-1 cells. A, a Venn diagram, summarizing microarray data analysis, shows the number of genes commonly regulated by Dex in THP-1 cells and in MΦ-THP-1 and uniquely regulated by Dex in both cell types. B, top 5 biological functions that are differentially regulated in THP-1 and MΦ-THP-1. C, heat map representing the top 25 up-regulated and the top 25 down-regulated genes by glucocorticoids in THP-1 and MΦ-THP-1 associated with the cellular movement pathway. D, in vitro transwell assay using 10% of FBS as chemoattractant was used to evaluate spontaneous cell migration of THP-1 and MΦ-THP-1 treated with vehicle, 100 nm Dex, 10 μm RU-486, or RU-486 with Dex. The graph shows that Dex treatment induces migration only in MΦ-THP-1, and this phenomenon is reversed by using the GR antagonist RU486. E, THP1-MΦ were transfected with NTC or GR siRNAs. 24 h after transfection, GR protein knockdown was evaluated by Western blotting (75% reduction). On the left, a representative immunoblot of GR and β-tubulin expression is shown. Right, densitometry analysis of GR normalized to β-tubulin. F, in vitro migration assay of THP1-MΦ transfected with NTC or GR siRNAs that have been treated for 24 h with or without 100 nm Dex. The histograms show that GR knockdown abolishes Dex-induced macrophage migration. Cell migration was calculated as percentage relative to vehicle-treated groups. Data are mean ± S.D. (error bars) and are representative of three independent experiments. ***, p < 0.001; ****, p < 0.0001; two-tailed unpaired Student's t test (E) and one-way ANOVA statistical test with Tukey's multiple-comparison test (D and F).