FIG. 6.

Differential effects of LPS on nuclear RARα protein levels in WT and IRAK-1^−/− BMDMs. (A) Effects of LPS on nuclear RARα levels in BMDMs. WT and IRAK-1^−/− BMDMs were treated with 100 ng/ml LPS for 2 h followed by nuclear protein extraction. The samples were analyzed by immunoblotting using the indicated antibodies. Lamin B was used as the loading control. (B) The BMDMs derived from WT and IRAK-1^−/− mice were treated with LPS for 2 h followed by RNA extraction. The resulting cDNAs were used to detect RARα transcript levels by real-time RT-PCR and standardized against GAPDH levels. (C) WT BMDMs were treated with 100 ng/ml LPS for 2 h followed by whole-cell protein extraction. The samples were resolved by SDS-polyacrylamide gel electrophoresis followed by immunoblotting with anti-RARα antibodies. (D) BMDM cells derived from WT and IRAK-1^−/− mice were either untreated or treated with leptomycin B (LepB) alone or in the presence of LPS. After 2 h of incubation, nuclear lysates were prepared and subjected to SDS-polyacrylamide gel electrophoresis followed by Western blot analysis with RARα-specific antibodies. The blots were also probed with lamin B-specific antibodies as a loading control.