Figure 7.

CD-MPR palmitoylation upon 19°C temperature block in vivo. (A) Mouse L cells stably expressing MPR wt and MPR-FFW-YLL-A were preincubated at 19 or 37°C for 30 min and then labeled with [³H]palmitate for 90 min at 19 or 37°C, respectively. The cells were then chilled on ice and lysed. CD-MPR was immunoprecipitated and subjected to SDS-PAGE. [³H]palmitate incorporation into CD-MPR was visualized by autoradiography. (B) Level of CD-MPR expression. Ten percent of the immunoprecipitated sample was subjected to SDS-PAGE and Western blotting (WB) with 22D4 mAb against CD-MPR. (C) Quantitation of [³H]palmitate incorporation into CD-MPR wt and FFWYLL-A. The fluorography and the Western blot shown in A and B, respectively, and those from additional experiments were quantitated by densitometric scanning. In each experiment, the values obtained for the [³H]palmitate incorporation were corrected for the CD-MPR expression levels. The value obtained with the CD-MPR wt incubated at 37°C was set to 1. The values are expressed as mean ± S.D. from three separate experiments.