Fig. 6

DAC induces AMPK degradation by increasing MAGEA6 expression and reducing MAGEA6 promoter hypermethylation. (A and B) RNA-seq analysis of DAC- and Ctrl-treated MOLM-13 cells. (C and D) MAGEA3 and A6 transcript levels were measured in MOLM-13 and MV4-11 cells treated with or without DAC (0.5 µM) for 48 h. (E) MAGEA6 protein level was measured in MOLM-13 and MV4-11 cells treated with DAC (0.5 µM) for 48 h. (F) Methylation-specific PCR (MSP) and unmethylation-specific PCR (UMSP) were performed to measure the methylation level of CpG island 1 in eight AML samples, six AML cell lines, and eight NCs. M: DNA Marker; B: Blank. (G) Bisulfite-genomic sequencing was performed to measure the methylation status of CpG island 1 in two NCs, two AML samples, and two AML cell lines. (H and I) MOLM-13 and MV4-11 cells were incubated with or without DAC (0.5 µM) for 48 h. DNA was extracted for bisulfite-genomic sequencing in Ctrl- or DAC-treated AML cells, and a summary of the frequencies of methylated CpG dinucleotides is shown. (J) The protein expression levels of MAGEA6, p-AMPK, AMPK, SLC7A11, and GPX4 were measured in MOLM-13 and MV4-11 cells transduced with pLVX-MAGEA6 overexpressing MAGEA6 (OE) or blank vector pLVX-NC (NC). *P < 0.05; **P < 0.01; ***P < 0.001 versus Ctrl cells. N.S: not significant