FIG. 4.

NFAT regulates ABCA1 expression. (A) Alignment of human and mouse sequences of the ABCA1 promoter. The mismatched nucleotides between the mouse and human sequences are highlighted in bold. The DR4 and the adjacent NFAT binding sites are indicated. (B) Direct binding of NFATc2 to the proximal promoter of ABCA1 in IRAK-1^−/− BMDMs. Cross-linked chromatin from untreated or ATRA-treated BMDMs (WT and IRAK-1^−/−) was incubated overnight in the presence of an antibody against NFATc2. Immunoprecipitated DNA was subjected to PCR amplification using primers flanking the potential NFAT binding sites on the ABCA1 promoter. The arrowhead points to the specific band. (C) Elevated levels of NFATc2 in the nuclear fraction of IRAK-1^−/− BMDMs. Untreated or ATRA-treated WT or IRAK-1^−/− BMDMs were subjected to nuclear fractionation followed by SDS-polyacrylamide gel electrophoresis and Western blot analysis using NFATc2-specific antibodies. The same blots were probed with lamin B-specific antibodies as a loading control.