Figure 2.

CD36 protein turnover is decreased in ob/ob macrophages. Pooled WT or ob/ob macrophages were pulse-labeled with [³⁵S]methionine/cysteine cell-labeling mix for 20 minutes (A) or 4 hours (B), then chased in medium with methionine/cysteine for the times indicated. ³⁵S-labeled CD36 in total lysates was immunoprecipitated with anti-CD36 and subjected to SDS-PAGE followed by transfer to nitrocellulose membrane and autoradiography. ³⁵S intensities were quantified by densitometric analysis, and each number was normalized for input (measured by Western analysis of β-actin in lysates), then to the relative intensity at chase time 0 hours. For 20-minute and 4-hour labeling experiments, one experiment representative of two and four independent experiments, respectively, is shown. Macrophages were isolated from five to seven mice of each strain.