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. 2024 Mar 22;300(5):107214. doi: 10.1016/j.jbc.2024.107214

Figure 2.

Figure 2

FADS1 supports AML cell survival and disease propagation in vivo.A, Western blot analysis of FACS-purified GFP+ MLL-AF9 cells expressing shNT, shFads1.1, or shFads1.2 (B) FACS-purified GFP+ MLL-AF9 cells from each shRNA condition were cultured in cytokine-enriched methylcellulose for 7 days (∗∗∗∗p < 0.0001). C, FACS-purified MLL-ENL cells from each shRNA condition were cultured in cytokine-enriched methylcellulose for 7 days (∗∗∗∗p < 0.0001). D, growth curve of Dnmt3aR878H; FLT3ITD-expressing mouse AML cells depicting fold change in % GFP+ cells over time (∗∗∗∗p < 0.0001 for all time points except day 7, shNT versus shFads1.2, ∗∗∗p = 0.0003). E, overall survival of mice transplanted with shNT, shFads1.1, or shFads1.2-expressing MLL-AF9 cells. Log-rank (Mantel-cox) test (n = 6 per group). F, Western blot analysis of FACS-purified GFP+ NOMO1 cells expressing shNT, i-shFADS1.1, or i-shFADS1.2, 48 h post-2 μg/ml doxycycline (DOX) treatment. G, cells from each inducible shRNA condition were treated with 2 μg/ml DOX and then counted at the indicated time points using flow cytometry counting beads on a LSRII flow cytometer (i-shNT versus i-shFADS1.1: ∗p = 0.0153, day 4 and ∗∗p = 0.0015, day 5; i-shNT versus i-shFADS1.1: ∗p = 0.0351, day 4 and ∗∗p = 0.0029, day 5). Dots represent individual data points and error bars represent SD. AML, acute myeloid leukemia; ENL, eleven nineteen protein; FACS, fluorescence-activated cell sorting; FADS1, fatty acid desaturase 1; MLL, mixed lineage leuekmia; shFADS1, shRNAs that reduce Fads1 protein expression; shNT, nontargeting control shRNA.