Research Paper Volume 12, Issue 17 pp 16759—16774

Figure 4. HDAC4 is required to maintain TET2 activity. (A, B) HDAC4 was overexpressed in 293T cells and TET2 expression determined by RT-qPCR (A) or Western blot (B). (C, D) After HDAC4 knockdown using lentivirus to deliver shRNA to MDS-L cells, 5hmC levels were determined by ELISA (C) or dot blot (D). (E) hMeDIP-qPCR analysis of specific 5hmC enrichment on Chr15: 58979469-58979554, enhancer region of MTSS1 in c-kit⁺ cells from BM of NHD13/HDAC4 WT or NHD13/HDAC4 KO mice. Bars represent mean enrichment over input. (F) MTSS1 mRNA levels in c-kit⁺ cells from BM of NHD13/HDAC4 WT or NHD13/HDAC4 KO mice. (G) RT-qPCR detection of MTSS1 mRNA levels in c-kit⁺ cells from BM of NHD13 mice treated with SAHA or vehicle control. (H) Serial replating of NHD13/HDAC4 WT or NHD13/HDAC4 KO cells. Total BM cells from each these mice were seeded in methylcellulose medium, and colonies were counted on day 7.