Anti lrp1

For SR-B1, LDLR, LRP1, SR-A1, and CD36 expression analyses, proteins of liver homogenates (50 μg total protein/lane) were separated by SDS-PAGE (8%) and transferred onto polyvinylidene difluoride membranes. The membranes were blocked with BSA (5%) and incubated with anti-SR-BI (1:1000, Novus), anti-LRP1 (1:500, SantaCruz), anti-LDLR (1:500, SantaCruz), anti-SR-AI (1:500, SantaCruz) and anti-CD36 antibodies (1:500, SantaCruz). This was followed by incubation with horseradish peroxidase (HRP) linked secondary IgGs (1:3000, SantaCruz) and visualization of the proteins by enhanced chemiluminescence (PerkinElmer).
To determine TNF formation by macrophages, THP-1 monocytes were stimulated with PMA to differentiate them into macrophages and then transfected with either control or Atg7-specific siRNA for 48 h. Transfected macrophages were then treated with either vehicle, LPS (100 ng/ml) or IFNγ (20 ng/ml) for 18 h. Macrophages were collected and lysed with Triton X-100 (0.4%). Subsequently, the lysate was centrifuged at 15,000 × g for 15 min; the supernatant was removed and kept on ice. Lysates (50 μg/lane) were separated by SDS-PAGE (10%) and transferred onto polyvinylidene difluoride membranes. After incubation with anti-TNFα antibody (1:1000, Genzyme), membranes were incubated overnight with HRP linked secondary IgG and the proteins were visualized by enhanced chemiluminescence.