Figure 1.

FOXC1 confers a differentiation block in human AML cells

(A–E) Human Fujioka AML cells were infected with a lentivirus targeting FOXC1 for KD or a non-targeting control (NTC). (A) Western blot shows FOXC1 KD 72 h after KD initiation. (B) Day 7 cytospins. (C) Bar chart (left panel) shows mean + SEM CD86 cell fluorescence (MCF) on day 5 (n = 3). Representative flow cytometry plots (right panel). (D) Bar chart (left panel) shows mean + SEM colony-forming cell (CFC) frequencies relative to control cells after 12 days in semi-solid culture (n = 3). Right panel: representative images. (E) Western blot.

(F–I) Primary patient AML cells (BB475) were infected with a lentivirus targeting FOXC1 for KD or a NTC with puromycin drug resistance as selectable marker (n = 2). (F) Western blot shows FOXC1 KD in BB475 AML cells 72 h following KD initiation. (G) Day 7 cytospins. (H) Bar chart (top panel) shows mean + SEM cell fluorescence (n = 2). Bottom panel: representative flow cytometry plots. (I) Bar chart (left panel) shows mean + SEM CFC frequencies of KD cells relative to control cells after 10 days in semi-solid culture (n = 2). Right panel: representative colonies. ^∗p < 0.05 for the indicated comparisons by t test (C, D, and I) or one-way ANOVA with Fisher’s least significant difference post hoc test (H).

Ap, apoptotic cell; Mφ, macrophage; MTV, empty vector; SDM3, site-directed mutagenesis construct #3. See also Figure S1.