FIGURE 9.

Localization of TM6SF2 and neutral lipids in mouse primary hepatocytes. A and B, primary hepatocytes from 8-week-old female WT mice that were fed chow ad lib were plated on coverslips for 4 h, fixed, and stained with BODIPY and with an antibody against TM6SF2 (A) and the LD marker PLIN2 (perilipin 2) (B). All images were taken using a ×63 oil immersion objective. Scale bar, 10 μm. C, female C57Bl/N mice (14 weeks old) were fed a high sucrose diet for 2 weeks. Feeding was synchronized for 3 days, and then the mice were killed at the end of the feeding cycle. Livers were homogenized, and the LDs, membranes, and cytosol were separated by ultracentrifugation as described under “Experimental Procedures.” Aliquots of proteins from the membrane and cytosolic fractions (50 μg each) and one-tenth of the LD protein was subjected to 10% SDS-PAGE and immunoblotting as described under “Experimental Procedures.” Calnexin (CNX), lactate dehydrogenase (LDH), and PLIN2 were used as controls for the ER, cytosolic, and LD fractions, respectively. The experiments were repeated, and the results were similar.