Knockdown of Utx augments the oncogenic activity in the murine T-ALL cell line MOHITO. (A) Western blot analysis of GFP sorted MOHITO knockdown samples containing 3 different hairpins against Utx compared with empty vector control. Blots were incubated with antibodies against Utx and tubulin. Expression levels were normalized to tubulin levels and compared with the vector control. (B) Graphical illustration of the interleukin depletion assay in the IL2-IL7–dependent MOHITO cell line. MOHITO cells are partially transduced with vectors encoding shRNAs against Utx and coexpressing GFP, after which the GFP percentage is measured by fluorescence-activated cell sorter analysis before and after successive rounds of interleukin depletion. (C) GFP percentage measured at Day 0, and after 1st and 2nd depletion rounds in Utx sh#1 and Utx sh#3 knockdown samples and controls. The depletion assay was repeated twice. The GFP enrichment was statistically significant after the 1st depletion for Utx sh#1* and Utx sh#3**, and after the 2nd depletion for Utx sh#1*** compared with empty vector control respectively (unpaired Student t test: *P = .005; **P = .049; ***P = .008).