Fig. 3: Targeting WNK1 prolongs survival of mice with AML.

a Schematic illustration of the experimental approach employed to generate tamoxifen-inducible Wnk1^fl/+, Wnk1^fl/− and Wnk1^fl/D368A MLL-AF9 leukaemia cell lines. c-KIT⁺ BM cells were transduced with MLL-AF9 oncogene and serially replated for five rounds in methylcellulose medium. The pre-leukaemia cells were transplanted into sublethally irradiated mice and leukaemia cells were isolated from the mice that developed leukaemia. b Growth curve of Wnk1^fl/+, Wnk1^fl/− and Wnk1^fl/D368A leukaemia cells in vitro treated with/without 500 nM OHT. Ethanol (EtOH) was used as a control. Data are presented as mean ± SD of three biological replicates (n = 3). c FACS analysis of EdU incorporation in Wnk1^fl/+, Wnk1^fl/− and Wnk1^fl/D368A leukaemia cells treated with 500 nM OHT for 48 and 72 h, showing a decreased S phase faction and an increased sub-G1 fraction, indicative of cell death. Data are representative of two independent experiments (n = 2). d Schematic of the experimental setup to test the requirement of WNK1 for AML in vivo. Sublethally irradiated B6.SJL recipient mice were reconstituted with Wnk1^fl/+, Wnk1^fl/− and Wnk1^fl/D368A leukaemia cells and the mice were treated with either tamoxifen or oil two weeks after transplantation. Mice were treated with Tamoxifen/oil for seven consecutive days and monitored for survival. e–g Kaplan–Meier survival curves of recipient mice transplanted with the indicated leukaemia cells. Statistical significance was calculated using a log-rank test.