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groups (Figure 7B). This indicated that Nsds-mediated H3K36me1/2 modifications were upstream of SEC com- plex recruitment and releasing promoter-proximal pausing of pol II. Meanwhile, SEC complex inhibitors could not affect other genes such as Gapdh and β-actin (Figure 7B).
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Besides the changes of protein expression levels, the functional changes were also assessed. The c-kit+ Setd2Δ/Δ cells were treated in vitro for 24 h with 3 inhibitors respec- tively, and LSK populations were further gated to analyze the apoptotic and cell cycle status. There were significant-
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Figure 5. Setd2Δ/Δ hematopoietic stem cells (HSCs) show loss of stem cell identity and increased differentiation toward progenitors. (A) Flow cytometry analysis of Setd2f/f and Setd2f/f/Vav1-Cre mice bone marrow (BM) cells. [N=3 each genotype; mean±Standard Error of Mean (SEM)]. (B) Gene Set Enrichment Analysis (GSEA) for genes affected in the LSKs of Setd2f/f and Setd2f/f/Vav1-Cre mice, after RNA-seq analyses. (A-E) Enrichment of HSC, long-term (LT)-HSC, short-term (ST)-HSC, early progenitors, intermediate progenitors, and late progenitors gene sets in LSKs of Setd2f/f and Setd2f/f/Vav1-Cre mice, respectively. All gene sets are from GSEA molec- ular signature database.26 (C and D) Up-regulated gene ontology analysis of differentially expressed genes. (E) Diagram of Setd2Δ/Δ HSPC differentiation.
haematologica | 2018; 103(7)