Fig 4.

IDOL induces late-endosome/lysosome localization of LDLR coincident with degradation. (A) HepG2 cells were treated with GW3965 (GW; 1 μM) or PCSK9 (5 μg/ml) for 1 h, and then proteasome inhibitor (25 μM MG132 [MG]) or lysosome inhibitor (50 nM bafilomycin [Baf]) was added for an additional 5 h. The levels of ABCA1 and LDLR were determined by immunoblotting. Similar results were obtained in three independent experiments. Numbers to the right of the blots are molecular weight markers. (B) HeLa cells were processed and analyzed as described for panel A. (C) HeLa cells were cultured in 10% LPDS medium for 8 h and then treated with GW3965 (1 μM) for the indicated times. Cells were immunostained with LDLR and LAMP-1 antibodies. Nuclei were counterstained with DAPI (blue). Representative confocal images are shown. T, time; O/N, overnight.