Effects of TIP110 on expression of CMYC and GATA2. (A) Total RNA was isolated from freshly isolated cord blood CD34⁺ cells, and the progeny of these cells were cultured overnight in IMDM medium supplied with 10% FBS and STF, and assessed by semiquantitative RT-PCR for TIP110, CMYC, and GATA2 gene expression. (B) Freshly isolated CD34⁺ cells were transfected with expression plasmids for GFP, TIP110.GFP, shRNA control (shCtrl), or shRNA for TIP110 (shTIP110). Cells were cultured in IMDM supplemented with FBS and STF (left) or without STF factors (right) for 48 hours. Total RNA was isolated by RT-PCR for TIP110, CMYC, and GATA2 expression. (C) NT-2 cells were transfected with shRNA control (shCtrl) or shRNA for TIP110 (shTIP110), cultured in DMEM supplemented with 10% FBS for 48 hours, and then harvested for TIP110, CMYC, and GATA2 expression by RT-PCR. (D) Human embryonic stem cells were transfected with shRNA control (shCtrl) or shRNA for TIP110 (shTIP110), cultured in HEScGRO hES cell medium for 72 hours, and then harvested for assay of TIP110, CMYC, and GATA2 expression by RT-PCR. Data for panels A through D are from 1 of at least 3 reproducible experiments.