AK2 knockdown induces early and massive apoptosis in T-ALL cell lines but not in B-cell ALL (B-ALL) cell lines. (A-B) Representative FACS graphs showing the percentage of Annexin V (AnnV) and propidium iodide (PI)-positive cells at day 2 (D2) and day 4 (D4) after shAK2 or shScramble transduction in JURKAT (A) and REH (B) cell lines. (C) Percentage of AnnV and PI-positive cells relative to control every 2 days from D2 post-shAK2 transduction to D8 in 5 T-ALL cell lines (red) and 2 B-ALL cell lines (blue). (D) Relative proliferation as a function of time. (E) Analysis of the different phases of the cell cycle (G₀/G₁, S, or G₂/M) at D2 in the shAK2 or shScramble condition after transduction in 5 T-ALL lines (JURKAT, RPMI-8402, PEER, MOLT4, and CEM) and 2 B-ALL lines (REH and RCH-ACV). (F) Relative proliferation to day 0 of five PDX T-ALL samples after transduction by shScramble (black) or shAK2 (red), measured with a CASY cell counter. (G) Percentage of AnnV and PI-positive cells at day 3 (D3) and day 7 (D7) performed on 5 PDXT-ALL samples after transduction by shScramble (black) or shAK2 (red). (H) Comparison of apoptosis induction between AK2-depleted PDX T-ALL (shAK2) and AK2-expressing cells (shsc) at D3 and D7 performed on 5 PDX T-ALL samples. All experiments were made in technical triplicates for cell lines and in technical duplicates for PDXs. *P < .05; **P < .01; ***P < .001. ns, not significant.