Fig. 6: The interaction of UHRF1 with SAP30 is critical for MXD4 repression and leukemogenesis.
From: Targeting UHRF1-SAP30-MXD4 axis for leukemia initiating cell eradication in myeloid leukemia
a The event free survival of AML patients was stratified by SAP30 expression into SAP30 high (survival days: 503 days, n = 211) and low (survival days: 646 days, n = 77) groups. b Correlation analysis of UHRF1/SAP30 expression in LSCs from AML patients was performed using the GSE76009 dataset. c The immunoprecipitation was performed using the anti-UHRF1 or anti-SAP30 antibody, and the anti-SAP30 and anti-UHRF1 antibodies were used for the Western blotting analysis in AML cells (n ≥ 3). d The GST pull-down assay shows that UHRF1 interacts with SAP30 in vitro and the SRA domain of UHRF1 is required for the interaction with SAP30. e The schematic representation of the truncations of SRA domain. f The immunoprecipitation assay was performed to examine the interaction of HA-tagged mutant UHRF1 with Flag-SAP30 in 293T cells (n = 3). g The number of colonies generated from UHRF1-deficient AML cells transduced with WT or mutant UHRF1 (n = 3). h The survival of B-NDG recipient mice receiving Kasumi-1 (n = 6) or THP-1 (n = 5) cells with the restoration of UHRF1 and UHRF1-Mut2 after UHRF1 knockdown. i, j MXD4 expression was examined by q-PCR (i) and Western blotting (j) analysis in AML cells with SAP30 knockdown (n = 3). k The CUT&Tag analysis shows that SAP30 binds to the promoter of MXD4 in AML cells. l The ChIP-qPCR analysis of UHRF1 on the promoter of MXD4 in AML cells with the knockdown of SAP30 (n = 3). m MXD4 expression was examined by q-PCR analysis in UHRF1-deficient AML cells transduced with WT or mutant UHRF1 (n = 3). n, o The representative DNA methylation profiles (n) and quantification (o) analysis of MXD4 by the bisulfite sequencing in G572R and F573R mutant AML cells with UHRF1 knockdown. Data are all presented as means ± SD. Statistical analyses were performed using log-rank test for a, h, and Pearson’s correlation analysis for b. Student’s unpaired t-test was used for g, i, l, m, o. *P < 0.05, **P < 0.01, ***P < 0.001.
