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Fig. 5 | Journal of Hematology & Oncology

Fig. 5

From: DNMT3A mutation leads to leukemic extramedullary infiltration mediated by TWIST1

Fig. 5

TWIST1 represents a downstream target of D3Amut. a Western blot of DNMT3A and EMT markers in acute leukemia cell lines OCI-AML3, THP-1, Kasumi-1, U937, and NB4. b Western blot presents the EMT markers in GFP-positive OCI-AML3 strains and those sorted from murine BMs and brains via GFP tags. c Real-time PCR of the mRNA levels of TWIST1 in AML patients’ primary BM cells with DNMT3A WT and DNMT3A R882H mutations. Bar graphs show the mean ± SD of ten and eight cases in WT and mutant groups, respectively. d Western blot shows the DNMT3A and TWIST1 in AML patients’ primary BM cells with DNMT3A WT and DNMT3A R882H mutations. e Left panel shows the Western blot analysis of TWIST1 and DNMT3A in sorted OCI-AML3 cells located in murine brain. Those cells are transiently transfected by scramble siRNA (si-CTL) or two different siRNAs targeting DNMT3A mRNA (si-DNMT3A-1 and 2). Transwell assays of OCI-AML3 cells from murine brain with or without DNMT3A mRNA knocked down are shown in the right panel. About 200 purified cells are used for plating in one well. Data are presented as mean ± SD for each group (n = 4). f Western blot of DNMT3A and TWIST1 in OCI-AML3 cells sorted from murine brain and are transiently transfected by si-CTL or siRNAs targeting TWIST1 mRNA (si-TWIST1 and si-TWIST2). Transwell assays of those sorted cells with si-CTL, si-TWIST1, or si-TWIST2. Cells are seeded in a number of 1 × 104. Data are expressed as mean ± SD (n = 6 per group)

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