TRIM13 overexpression represses leukemogenesis. (A) Western blot confirmation of TRIM13 overexpression or knockdown in AML cell lines. (B) In vitro suspension culture competitive growth assay. Values shown are mean of 3 independent replicates. (C) Colony formation assay in indicated AML cell lines expressing empty vector or TRIM13 overexpression. (D) Cell death assay measuring annexin V and propidium iodide staining in AML cell lines expressing empty vector of TRIM13 overexpression, taken at day 5. (E) Western blot confirmation of CHAF1B knockdown by shRNA. (F) CD14 surface marker expression on U937 cells 72-hours after CHAF1B knockdown. (G) CD14 surface marker expression on U937 and MOLM13 AML cell lines 5-days after TRIM13 overexpression. Red star indicates significant increase in CD14 expression when compared with empty vector. (H) Western blot confirmation of TRIM13 overexpression in sample of patients with AML. (I) Colony formation assay in indicated sample of patients with AML expressing empty vector or TRIM13 overexpression. (J) Schematic for xenograft transplantation of AML cell lines into NSG mice. (K) Western blot confirmation of TRIM13 overexpression in MOLM13 AML cell line. (L) Percent human CD45+ mononuclear cells in BM aspirates 26 days after transplantation. Each dot represents an individual mouse (n = 10), and line indicates mean. (M) Survival curve of recipient mice receiving TRIM13 overexpressing or control AML cell lines (n = 10 mice). ∗P < .05, as determined using nonparametric t test with Welch correction (C,D,L), one-way ANOVA (I), and log-rank test (M). Data shown are representative of 3 independent assays (A,E-H,K) and the mean of 3 biological replicates (B) with SD (C-D). FACS, fluorescence-activated cell sorting.