Figure 7: PRC1 contributes to development of DNMT3A mutant-associated leukaemic cells.

(a) Design of CFU assay and liquid culture assay with HOXA9 and EV-, WT- or R882H-transduced c-kit+ Bmi1^+/+ or Bmi1^+/− murine BM cells. (b) Growth curve of Bmi1^+/+ and Bmi1^+/− murine immortalized cells transduced with HOXA9 and EV or R882H in liquid culture; N=3 replicate experiments. (c) Typical morphology of Bmi1^+/+ and Bmi1^+/− murine immortalized cells transduced with HOXA9 and EV or R882H in liquid culture. Scale bar, 20 μm. (d) Representative surface-marker profile of Bmi1^+/+ murine immortalized cells transduced with HOXA9 and R882H. (e) Cumulative colony-forming capacity is shown for an initial plating of 1,000 co-transgene-expressing cells; N=3 replicate plates. P values were calculated using two-tailed unpaired t-test. (f) Relative expression levels of PU.1 in co-transduced cells at first round measured by qPCR; N=3 replicate experiments, normalized to the expression levels in HOXA9 and EV co-transduced cells. P values were calculated using two-tailed unpaired t-test. BM, bone marrow; CFU, colony-forming unit; EV, empty vector; PRC1, polycomb repressive complex1; qPCR, quantitative polymerase chain reaction; WT, wild type. Data are mean±s.d. values. *P<0.05, ^**P<0.01.