Skip to main content
. 2021 Jun 3;12(6):573. doi: 10.1038/s41419-021-03738-0

Fig. 1. KDM4A is required for the functional potential of human and murine AML cells.

Fig. 1

A–E Human THP1 AML cells were transduced with lentiviruses targeting KDM4A or other KDM4 family members for KD (♯1 and ♯2 represent individual distinct lentiviruses targeting genes for KD as indicated), or a non-targeting control (NTC), using KD of MLL and MEN1, as positive controls, which are essential genes for AML cell proliferation. All bar charts show mean ± s.e.m. A Resorufin signal after 4 days of individual KDM4 family member KD relative to NTC control cells (n = 3); *p < 0.01 for comparison of each KD versus NTC. B Expression of KDM4A/B/C/D in indicated KD cells relative to NTC control cells (n = 3); *p < 0.001. C Representative immunoblot showing KDM4A KD in THP1 cells (n = 3). D Scatter plot shows the correlation of KDM4A KD with inhibition of frequency of colony-forming cells (CFC) enumerated following 10 days in semisolid culture (n = 3), as determined by QPCR; *p < 0.001. E Percentage of apoptotic cells determined by Annexin V+/ 7AAD+/− staining on day 4 of liquid culture after puromycin selection (n = 3); *p < 0.001. F–G The indicated primary unfractioned patient blasts were transduced with lentiviruses targeting KDM4A for KD, or an NTC. Primary AML cells used include BB160, containing t(9;11) (MLL-AF9) chromosomal translocation and BB86 (normal cytogenetics, non-MLL) (BB number is the Manchester Cancer Research Centre Biobank sample identifier). All bar charts show mean ± s.e.m. F CFC frequencies of primary human AML blasts (n = 3) following lentivirus infection, puromycin selection, and initiation of KDM4A KD; *p < 0.0001. G Representative images from F. H CFC frequencies of primary murine MLL-AF9 AML cells following KDM4A depletion (n = 3); *p < 0.0001. I Survival curves of NSG mice transplanted with 10,000 KDM4A KD or NTC THP1 cells (n = 5 per cohort); p by log-rank test. J Survival curves of NSG mice transplanted with 106 KDM4A KD or NTC primary AML cells (BB160, n = 7 per cohort); p by log-rank test. K Spleen weights of mice from J with a representative image of the spleen. p by one-way ANOVA, F = 34.13045.