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. 2023 Nov 9;109(4):1082–1094. doi: 10.3324/haematol.2023.283437

Figure 5.

Figure 5.

Extended-dose Aza upregulates myeloperoxidase expression to induce leukemic stem cell differentiation, which can be partly rescued by myeloperoxidase inhibition or reactive oxygen species scavenging. (A) OCI-AML20 cells were sorted to retrieve the CD34+38- population. (B) Sorted cells were treated with azacitidine (Aza) (extended or conventional dose) and bulk RNA sequencing was performed at day 5 followed by pathway enrichment analysis (gene ontology [GO] process) to assess Aza-induced pathways. (C) OCI-AML20 cells were treated with Aza (conventional or extended regimen) and myeloperoxidase (MPO) expression was assessed by capillary electrophoresis on day 7, glyceraldehyde-3-phos phate dehydrogenase (GAPDH) was used as a loading control. (D) Representative flow data from 1 experiment with OCI-AML20 cells were treated with extended or conventional regimen of Aza with or without 10 µM MPO-IN-28 or (E) N-acetylcysteine (NAC) on days 2 and 4 and flow cytometry assessment was conducted on day 7 using live/dead, CD45, CD34 and CD38 staining. DMSO: dimethyl sulfoxide.