Research Paper Volume 12, Issue 17 pp 16759—16774

Figure 1. SAHA treatment reduces TET2 and 5hmC levels in HSPCs of NHD13 mice. (A) NHD13 mice were divided into two groups of matched gender and age. Peripheral blood (PB) was sampled before treatment, and mice were then treated 4 weeks with SAHA (50mg/kg/day, I.P., n = 8) or vehicle (5% DMSO in 20% cyclodextrin, I.P., n = 8). After treatment, mice were euthanized, and PB and BM were collected. (B) CBC as indicated in NHD13 mice analyzed after 4 weeks of treatment. (C) Hematopoietic lineage subset frequencies in BM of NHD13 mice after 4 weeks of indicated treatment. (D) Hematopoietic progenitor subset frequencies in BM of NHD13 mice after 4 weeks of indicated treatment. (E, F) 5hmC levels in MNCs of PB from NHD13 mice before or after treatment, as determined by ELISA (E) and dot blot (F). Methylene blue staining served as a loading control. 5hmC levels were normalized against loading of total DNA. (G) 5hmC levels in LSK cells of BM from treated NHD13 mice (n = 3), as determined by intracellular staining with a 5hmC-specific antibody. (H) Western blot showing TET2 protein levels in c-kit+ cells from BM of NHD13 mice treated as indicated. (I) RT-qPCR detection of TET2 mRNA levels in c-kit+ cells from BM of NHD13 mice treated as indicated.