Figure 6
From: Phospholipase D1 deficiency in mice causes nonalcoholic fatty liver disease via an autophagy defect
Hepatic PLD1 expression in Pld1−/− mice attenuates hepatic steatosis.
(a) Primary hepatocytes were isolated from Pld1+/+ and Pld1−/− littermates, infected with adGFP or adPLD1 for 24 h, and treated with or without serum for 24 h. Western blot was performed with cell lysates. (b) Primary hepatocytes were infected with adGFP or adPld1 for 24 h, serum starved, and then treated with bovine serum albumin/palmitate complexes for 2 h. The OCR was measured at the indicated time points. (c) After feeding the mice with a HFD for one week beginning at 13 weeks, the mice were injected with adGFP or adPld1 (1 × 109 PFU) by tail vein injection. At 12 days post-injection, mice were sacrificed for analyses. Mouse liver lysates were analyzed by western blotting. (d) Mice were maintained as described in (c) and the livers were isolated, and frozen sections were stained with H&E (left) and ORO (right). Representative images were obtained at 200× magnification from 3–5 independent experiments. Scale bar, 50 μm. (e) Mice were maintained as described in (c); the weights of livers from mice were measured and hepatic triglycerides and cholesterol were quantitatively analyzed. The data are presented as means ± SE or representative blots from 3 to 5 independent experiments. *P < 0.05 versus Pld1+/+ mice. Abbreviations: adGFP, adenovirus-expressing green fluorescence protein; adPld1, adenovirus-expressing phospholipase D1; H&E, hematoxylin and eosin; HFD,; OCR, oxygen consumption rate; PA, palmitic acid; PFU, plaque-forming units; SE, standard error.
