Fig. 3: Levels of TET2 expression and activity regulates proliferation in human myeloid cells.

a Natural TET2-mutant HEL and TET2 wild-type MEG-01 and CMK cells were stably transfected with either an empty pOZ (Vector) or a pOZ-TET2-overexpression vector (TET2^OE). Levels of 5hmC and 5mC were assessed by dot blot assay. b–d Cell lines (Vector and TET2^OE of HEL, MEG-01, and CMK) were treated for 6 days with either PBS or 250 µM AA. Surviving cells were assessed by trypan blue exclusion assay on Vi-CELL XR cell viability analyzer (Beckman Coulter) and the cell output was plotted. b MEG-01. c CMK. d HEL. e–h Human K562 and MOLM13 cells were transduced with scrambled shRNA (scr) or shRNA targeting TET2 (shTET2) and treated with 250 µM AA. e–f 5hmC and 5mC were assessed after 24 h by 2D-UPLC-MS/MS. g–h Cellular proliferation was accessed by the number of viable cells at different time. i–j Primary human cord blood CD34⁺ cells and proliferation was assessed after 14 days by colony forming assay. a–j Data are shown as mean ± SEM (n = 3) and are representative of two independent experiments. k TET2^WT (n = 8) and TET2^MT (n = 11) human BM levels of 5hmC/5mC was measured by 2D-UPLC-MS/MS. Data are shown as mean with SEM. l TET2^WT and TET2^MT human BM cells were treated for 48 h with 250 µM AA or left untreated. Base oxidation levels were assessed by 2D-UPLC-MS/MS. Data from the same cell are connected with arrow, TET2^WT, n = 3; TET2^MT, n = 4. TET2^MT MN patient information are provided in Supplementary Data 4. Paired t test was used in (l), p values are indicated, ns: not significant.