CCL5-inducible up-regulation of cyclin D1 and MMP-9 protein levels is dependent on mTOR-mediated mRNA translation. (A) Activated PB T cells were either pretreated with DMSO (carrier) or 50 nM rapamycin for 1 hour prior to treatment with 10 nM CCL5 for the indicated times. Cells were harvested and lysates resolved by SDS-PAGE and immunoblotted with anti–cyclin D1 or anti–MMP-9 antibody. Membranes were stripped and reprobed for β-tubulin as loading control. The relative fold increase of cyclin D1 protein level is shown as signal intensity over loading control. Data are representative of 3 independent experiments plus or minus SD. (B) T cells were either left untreated or treated with 10 nM CCL5 for 1 hour and the mRNAs extracted. Semiquantitative RT-PCRs were performed using primer sets specific for cyclin D1, MMP-9, and GAPDH, as described in “Methods.” Data are representative of 2 independent experiments.