CHAF1B upregulation and TRIM13 downregulation are associated with AML progression. (A-B) Schematic representation of Chaf1b-floxed allele in mouse MLL-AF9 leukemic cells and confirmation of Chaf1b deletion after Cre activation via quantitative polymerase chain reaction (qPCR) (A) and western blot (B). Value shown is mean ± standard deviation (SD) of n = 3 independent experiments. (C) Western blot at 24 and 48 hours after Cre induction in MLL-AF9 leukemic cells. (D) Survival of patients with AML with high or low TRIM13 expression in the Cancer Genome Atlas (TCGA). P value shown. (E) Expression of TRIM13 throughout human hematopoiesis and leukemia from BloodSpot. (F) Scatterplot of expression comparing CHAF1B and TRIM13 in TCGA. The solid red line represents best fit with ± SD (dashed red lines). Line equation, P value, and R² value listed. (G) Immunohistochemistry (IHC) in BM biopsies for CHAF1B (top), TRIM13 (middle), and hematoxylin and eosin (H&E) staining of biopsies of healthy BM and from 3 matched patients with initial/relapse or initial/remission AML. Scale bar, 20 μm; IHC and H&E imaged from the same section, if possible. (H) Western blot confirming TRIM13 short hairpin RNA (shRNA) efficacy in mobilized CD34⁺ PBSCs. Results shown are representative of 3 independent assays. (I) Western blot confirming the CHAF1B overexpression efficacy. Results shown are representative of 3 independent assays. (J) Colony assay in PBSCs expressing indicated viral constructs. Results shown are mean ± SD, dots indicate individual replicates. ∗P < .05, as determined using 1-way analysis of variance (ANOVA) (J). SCRsh, scrambled control short hairpin; Veh, vehicle.