HA117 endows HL60 cells with a stem-like signature by inhibiting the degradation of DNMT1 via its ability to down-regulate expression of the GGL domain of RGS6
Fig 3
HA117 down-regulated the expression of the GGL domain of RGS6.
HL60/ATRA and generation-matched wild HL60 cells were cultured in growth medium (RPIM1640 with 10% FBS, CTR) and harvested during logarithmic growth phase. Cells were then subjected to (a) real-time PCR analysis and (b) western blot analysis. HL60/ATRA cells were infected with empty lentiviral vectors (HA-con) or HA117-specific shRNAs (HA-si), and the infected cells were selected with puromycin for 2 days. These cells were amplified and harvested, and subjected to (c) western blot analysis and (d) real-time PCR analysis. Transfection of HA117 shRNA up-regulated GGL domain expression in HA-si cells. HA-si cells were infected with lentiviral vectors carrying control scrambled (HA-si-con) or full-length HA117 (HA-si+117), followed by puromycin selection of the infected cells for 2 days. Selected cells were then expanded and harvested, and subjected to (e) western blot analysis and (f) real-time PCR analysis. Transfection of full-length HA117 suppressed GGL domain expression in HA-si+117 cells.*P<0.005 (Student’s t-test). All statistical analyses were performed with Microsoft Office Excel.