Figure 3.

PTEN knockdown confers a selective advantage to diagnosis leukemia cells. (A) Schematic of the protocol for competitive engraftment experiments shown in B and C. Case XLE_61 T-ALL diagnosis cells transduced with control shRNA (shRNA CTL and CTL-Cherry⁺ cells) and shRNA PTEN (PTEN^KD-GFP⁺ cells) lentivectors were sorted and co-transplanted in equal number into NSG mice. Two cell doses (5 × 10⁴ and 5 × 10³ cells per mouse) were tested; three to five mice were injected per cell dose. Immunoblots show PTEN silencing efficiency. (B) Flow cytometry analyses showing the proportion of PTEN^KD-GFP⁺ and CTL-Cherry⁺ cells in leukemia cells immediately before transplantation (left) and in BM cells of a representative mouse after establishment of leukemia (right). (C) PTEN^KD-GFP⁺ versus CTL-Cherry⁺ CD45⁺ cells ratios are indicated, and each dot represents data of one mouse. The dotted line indicates ratio 1. (D) Kinetics of engraftment of cells transduced with shRNA CTL (CTL-GFP⁺ cells) and shRNA PTEN (PTEN^KD-GFP⁺ cells) lentivectors, injected in parallel (noncompetitive) experiments with decreasing cell doses, as measured by the percentage of CD45⁺GFP⁺ cells in BM cells. Each dot represents data of one mouse, dashes represent medians, and lines connect the medians.