Functional cooperation of NFIA-ETO2 with TP53^R248Q in vitro. (A,D,G,J) Growth curves of TP53^R248Q/+ (A), Tp53^+/− (D), TP53^R248Q+/− (G), and Tp53^−/− (J) BM-derived EBs expressing NFIA-ETO2 (red line) compared with CTRL cells (blue line) over 6 days in DM (n = 2-3). (B,E,H,K) Representative images of Wright-Giemsa-stained cytospin preparations (top) and flow cytometry panels showing CD71/Ter119 expression (bottom) of TP53^R248Q/+ (B), Tp53^+/− (E), TP53^R248Q/− (H), and Tp53^−/− (K) BM-derived EBs expressing NFIA-ETO2 compared with CTRL grown in MM (left) and in DM (right). Data represent 1 out of 2 to 3 independent experiments. Images were recorded with a 60× objective using a Nikon-TI. Scalebars, 50 μm. (C,F,I,L) Colony formation in MC by TP53^R248Q/+ (C), Tp53^+/− (F), TP53^R248Q/− (I), and Tp53^−/− (L) BM-derived EBs expressing NFIA-ETO2 compared with CTRL. Shown are absolute numbers of colonies formed in 6 consecutive platings (n = 2-3). Note that control cells cannot self-renew after 2 rounds of plating. Values are presented as individual points, bar graphs represent the mean value of independent biological replicates (n); and error bars are standard error of the mean. Statistical significance was tested with paired two-tailed t tests.