Figure 1

Acute alcohol represses expression and promoter activity of MFG-E8 gene in macrophages. (A and B) Effect of acute alcohol exposure on MFG-E8 gene expression. Murine macrophages were treated with culture medium alone (control) or medium containing alcohol (25 mmol/L) for 18 h as indicated in the figure. At the end of treatments, total cellular RNA and protein were extracted from cells. MFG-E8 mRNA levels (panel A) in total RNA samples were quantified with real-time RT-PCR, whereas MFG-E8 protein (panel B) in total cellular protein samples were determined with Western blot as described in Materials and Methods. (C) Scheme of murine MFG-E8 gene promoter/luciferase reporter. A fragment of murine MFG-E8 gene promoter (−444 to +146) was cloned into the upstream of the luciferase gene in pGL3-basic reporter plasmid. The construct is named as pGL3-mfge8^−444/+146. (D) Acute alcohol represses MFG-E8 promoter activity. RAW 264.7 cells were dual transfected with plasmids of pRL-null and pGL3-mfge8^−444/+146 reporters. They were subjected to treatment with medium alone (control) or alcohol (25 mmol/L) for 18 h followed by processing for luciferase assay as described in Materials and Methods. The experiments were performed twice with triplicates. Results are expressed as mean ± SEM; n = 3. *P < 0.05 versus control group.