Fig 1.

The LXR-IDOL pathway blocks LDL association with LDLR at the plasma membrane in living cells. (A) HepG2 cells stably overexpressing GFP-LDLR were treated with GW3965 (1 μM) overnight and then incubated with DiI-LDL. The association of DiI-LDL was determined in living cells by spinning-disc confocal microscopy. Still images at time zero and 2.5 h are shown. The entire movie shown in Video S1 in the supplemental material covers 3 h. (B) Immunoblot analysis of IDOL protein expression in transiently transfected 293T cells with monoclonal antibody (10E7F10). Cells were treated with or without MG132 for 4 h prior to harvest. (C) Immunoblot analysis of endogenous IDOL protein expression with antibody 10E7F10 in wild-type (WT) and IDOL-deficient cells. Primary mouse peritoneal macrophages were starved in lipoprotein-deficient medium and then treated with GW3965 overnight and MG132 for 5 h. T, time.