Rapamycin enhances LV transduction via an mTOR-dependent, nonautophagy mechanism. (A) Human cord blood CD34⁺ cells were transduced with CG-UbiC-EGFP, MOI = 25, in the absence or presence of the autophagy-stimulating peptide Tat-beclin 1. Black triangles, Tat-beclin 1 scrambled peptide control; blue circles, Tat-beclin 1. Data are pooled from 2 independent experiments, each with different cord blood donors and done in duplicate. (B) Bone marrow Lin⁻ cells from wild-type or beclin 1^+/− mice were transduced with RRL-MND-GFP, MOI = 0.5, in the presence of 5 μg/mL rapamycin or DMSO. EGFP marking was assessed by flow cytometry 10 days posttransduction. Data points represent cells from individual donor mice. Black triangles, wild-type cells; blue circles, beclin 1^+/− cells. (C) Human cord blood CD34⁺ cells were transduced with CG-UbiC-EGFP, MOI = 25, in the presence of combinations of either rapamycin or (D) Torin 1 with FK506, an FKBP12-binding compound. Black triangles, DMSO only; green squares, 10 μg/mL FK506. Data are duplicate transductions from 2 representative experiments with different cord blood donors. For all panels, lines represent group mean and error bars represent standard deviation. ****P < .0001 from a parametric 2-tailed unpaired Student t test.