Journal: Journal of Lipid Research

Article Title: Regulation of SR-BI-mediated selective lipid uptake in Chinese hamster ovary-derived cells by protein kinase signaling pathways

doi: 10.1194/jlr.m600326-jlr200

Figure Lengend Snippet: Fig. 5. HDL, but not AcLDL, stimulates phosphorylation of the protein kinase C (PKC) substrate myristoylated alanine-rich C- kinase substrate (MARCKS). mSR-BI-overexpressing (ldlA[mSR- BI]) cells, prepared as described in the legend to Fig. 4, were given serum-free Hams F12 medium containing either no additions (lane 1), 0.5 mM PMA, HDL, or AcLDL (each at 50 mg/ml), 5 mM RO31-8220, a 1:1000 dilution of an anti-SR-BI blocking antiserum, or combinations as indicated. Cells were incubated for 3 min, at which time they were rapidly chilled on ice, then washed, lysed, and analyzed by SDS-PAGE and immunoblotting for either phospho- MARCKS or b-actin. The data are representative of multiple experi- ments. For the experiment shown, lanes 9 and 10 were from a different portion of the same gel.

Article Snippet: Rabbit anti-SR-BI antibody (NB400-101) and anti-phospho-myristoylated alanine-rich C-kinase substrate (MARCKS) (NB500-140) were from Novus Biologicals, Inc. (distributed by Cedarlane Laboratories Ltd, Hornby, Ontario, Canada).

Techniques: Phospho-proteomics, Blocking Assay, Incubation, SDS Page, Western Blot