Reduced expression of TLKs suppresses in vitro and in vivo AML growth and leukemia burden using human cells. (A) A schematic representation of a doxycycline-inducible shRNA model targeting TLK1, TLK2, or both TLK1 and TLK2 in MOLM-14 cells. (B) MOLM-14 cells were treated with 1 μg/mL doxycycline for 6 days to induce knockdown. The effect of the knockdown was measured by immunoblotting analysis. (C) Cell viability of MOLM-14 cells expressing shTLK1, shTLK2, or both was measured by colorimetric (MTS) assay after 6 days of doxycycline-induced knockdown. (D) Cell growth over time of MOLM-14 cells expressing shTLK1, shTLK2, or both was indicated by cell counts. (E) Cell cycle analysis of MOLM-14 cells expressing shTLK1, shTLK2, or both using propidium iodide and flow cytometry after 6 days of doxycycline-induced knockdown. (F) Colony formation was assessed by plating cells in methylcellulose-based medium after doxycycline-induced knockdown for 14 days. Data are expressed as means ± SEM. ∗P < .05, ∗∗P < .01, and ∗∗∗P < .001.