Figure 4.
TRIM13 repression promotes leukemogenesis. (A) Western blot confirmation of TRIM13 knockdown in PDX 16 to 35 before transplant. (B) Percent human CD45+ mononuclear cells in BM aspirates on indicated days after transplantation. Each dot represents an individual mouse (n = 10), middle line represents mean, box represents SD, and whiskers represent minimum-maximum range. Individual points represent individual mice (n = 10). (C) Survival of recipient mice after transplantation of 1 × 106 live AML16 to AML35 cells. (D) Western blot confirmation of TRIM13 knockdown in AML17 to AML129. (E) Colony forming assay in indicated PDXs expressing TRIM13sh2 (T13sh2). (F) Schematic diagram of RING domain deletion in TRIM13 by CRISPR. (G) Western blot confirmation of deletion clones in AML cell lines. ∗ indicates full-length TRIM13. (H) Colony assay in indicated AML cell lines comparing CTRLsg with T13RINGdel cells. (I) Cell death assay at steady state in T13RINGdel clones compared with CTRLsg. Red star indicates significant difference in AnnexinV/propidium iodide between conditions, black start indicates significant differences in AnnexinV between conditions. (J) Cell surface CD14 expression at steady state in AML cell lines. Unless otherwise noted, ∗P < .05, as determined using t test with Welch correction (B), one-way ANOVA (E,H-I), or log-rank (C). All experiments shown are representative of 3 biological replicates (A,D,G,J), mean + SD of 3 biological replicates (E,H,I), or from 10 mice (B-C). CTRLsg, cells electroporated with a nontargeting control single guide RNA.