Figure 3.

Induction of SREBP-1c transcripts in McA-RH7777 cells after treatment with LXR ligands. On day 0, cells were set up as described in Fig. 1. On day 2, the cells were washed with PBS and switched to medium B with (lanes 6–10) or without (lanes 1–5) 50 μM sodium compactin and 50 μM sodium mevalonate. The indicated sterol or the LXR-agonist T0901317 was added at a final concentration of 10 μM. After incubation for 16 h at 37°C, cells were harvested, and aliquots of total RNA (20 μg) from pooled dishes were hybridized for 10 min at 68°C to ³²P-labeled cRNA probes for rat SREBP-1, SREBP-2, and β-actin. Protected fragments were separated by gel electrophoresis and exposed to film for 8 h at −80°C as described in Materials and Methods. 25-HC, 25-hydroxycholesterol; 22(R)-HC, 22(R)-hydroxycholesterol.