Fig. 1.

Hepatocytes efficiently lipidate endogenous and exogenous SAA. A, C, D: SAA content of conditioned media and cell lysates from primary hepatocyte cultures. Mouse primary hepatocytes were prepared as described in the Materials and Methods and, after overnight culture, were treated with 5 μM T0901317 to induce ABCA1 expression, and then cultured in medium containing 0.2% fatty acid-free BSA. Conditioned media (7 μl) and cell lysates (10 μg protein) were collected at the indicated times were separated by SDS-PAGE followed by Western blotting to detect SAA with β-actin as loading control for cell lysates. A: Hepatocytes were prepared from untreated SAA1.1/2.1/3-TKO mice and C57BL/6 (C57) mice 6 h after injection of 1 μg/g body weight LPS. The results shown are representative of three immunoblots. B: Cell media from A, C, and D were subjected to NDGGE followed by Western blotting to detect SAA. The migration of size standards and lipidated SAA are indicated. The results shown are representative of two separate immunoblots. Note that a lower exposure of this immunoblot is presented in supplemental Fig. S1. C: Hepatocytes were prepared from untreated SAA1.1/2.1/3-TKO mice or SAA1.1/2.1/3-TKO mice 24 h after administration of 1 × 10¹¹ AdSAA. The results are representative of three immunoblots. D: Hepatocytes from untreated SAA1.1/2.1/3-TKO mice were incubated without or with 10 μg/ml purified mouse SAA; the results are representative of three immunoblots.