Figure 4. RELA is required for MLL leukemia development.

(A-C) Mouse MLL-AF9 cells were transduced with lentiviral vectors expressing control or Rela shRNAs. Protein levels of Rela were detected by western blot analysis (A). Cell numbers (B) and colony numbers (C) were enumerated after 2 and 5 days, respectively, and expressed relative to the numbers obtained with control shRNA transduced cells.

(D and E) Mouse MLL-AF9 cells were transduced with Rela over-expression or control vectors. Cell numbers (D) and colony numbers (E) were enumerated after 3 and 5 days, respectively, and expressed relative to the numbers obtained with control vector transduced cells.

(F) Hematopoietic progenitors obtained from mouse fetal livers (E13.5) of wild-type (Rela^+/+) or Rela knockout (Rela^-/-) embryos were transduced with the indicated oncogenes and used for serial myeloid replating assays. Representative results from two independent replicates through four rounds of serial methylcellulose culture are shown as colony number per 3000 cells.

(G) Survival curves are shown for cohorts of mice transplanted with mouse MLL-AF10 leukemia cells (5 × 10⁵) transduced with control or Rela shRNAs (n = 5 each cohort). Acute leukemia was confirmed by peripheral blood leukocyte count and necropsy. Log-rank Test was used for statistical analysis (p = 0.006).

(H) Survival curves are shown for mice transplanted with mouse MLL-AF6 transformed cells (1 × 10⁶) co-transduced with Rela or control vectors (n = 5 each cohort). Log-rank Test was used for statistical analysis (p = 0.002).

All error bars represent SD of triplicate analyses. See also Figure S4.