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Fig. 6 | BMC Cancer

Fig. 6

From: A novel miR-375-HOXB3-CDCA3/DNMT3B regulatory circuitry contributes to leukemogenesis in acute myeloid leukemia

Fig. 6

HOXB3 enhances the expression of DNMT3B to bind in pre-miR-375 promoter. a HL-60 and THP1 cells were transduced with special shRNA for HOXB3 (sh-HOXB3) or sh-NC. HOXB3 and DNMT3B expressions were detected by western blot. b HOXB3 and DNMT3B expressions were detected in HL-60 and THP1 cells, which were transduced with overexpression vector LVX-HOXB3 or LVX-NC. c DNMT3B expression was detected in HL-60 and THP1 cells transduced with special shRNA targeting DNMT3B (sh-DNMT3B) or sh-NC. d MiR-375 expression was measured in HL-60 and THP1 cells transduced with sh-DNMT3B or sh-NC. *P < 0.01 versus sh-NC. e Soluble chromatin from HL-60 and THP1 cells, which were transduced with sh-NC or sh-DNMT3B, was immunoprecipitated with anti-DNMT3B antibody. Immunoprecipitated DNA was analyzed by qRT-PCR. *P < 0.01 versus sh-NC. f Bisulfite genomic sequencing was performed to detect the methylation status of the DNA sequences at -260 bp − + 136 bp in the pre-miR-375 gene upstream region in HL-60 and THP1 cells, which were transduced with sh-NC or sh-DNMT3B. Each row of circles represents the sequence of an individual clone. Black circles and empty circles represent methylated and unmethylated CpG dinucleotides, respectively (Left). Shown was the summary of frequencies of methylated CpG dinucleotides detected in HL-60 and THP1 cells by bisulfite genomic sequencing (Right). *P < 0.01 versus sh-NC. g Bisulfite sequencing was performed using DNA from HL-60 and THP1 cells transduced with MSCV-DNMT3B or MSCV-NC. Black circles and empty circles represent methylated and unmethylated CpG dinucleotides, respectively (Left). Shown was the summary of frequencies of methylated CpG dinucleotides detected in HL-60 and THP1cells by bisulfite genomic sequencing (Right). *P < 0.01 and #P < 0.05 versus MSCV-NC. h Overexpression of DNMT3B in HL-60 and THP1 cells was indicated by western blot

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