Figure 1
Comparison of 2i treatment on UHRF1 and DNMT1 expression in mESCs and HCT116 cells. (a) Western blotting analysis showing 2i down-regulated the levels of UHRF1 and DNMT1 proteins in mouse embryonic E14 cells. E14 cells were maintained in serum plus LIF with or without addition of 2i and subjected to western blotting analysis using antibodies as indicated. (b) Addition of MG132 blocked 2i-induced down-regulation of UHRF1. E14 cells maintained in serum plus LIF or 2i medium were treated with MG132 for 6 hours before harvested for western blotting analysis. (c) qRT-PCR analysis showing the 2i down-regulated the mRNA levels of DNMT3A, but not UHRF1 and DNMT1, in mouse embryonic E14 cells. E14 cells were maintained either in serum plus LIF or 2i medium and subjected to qRT-PCR analysis. (d) 2i also down-regulated UHRF1 and DNMT1 but not DNMT3A in HCT116 cells. The HCT116 cells were treated with or without addition of 2i for 2 days before harvested for western blotting analysis. (e) Addition of MG132 could not block 2i-induced down-regulation of UHRF1 and DNMT1 in HCT116 cells. The HCT116 cells were treated with or without addition of 2i for 2 days and MG132 was added 12 hours before cells were harvested. Western blotting analysis was performed using antibodies as indicated. (f) qRT-PCR analysis showing 2i down-regulated the mRNA levels of UHRF1 and DNMT1, but not DNMT3A, in HCT116 cells. The HCT116 cells treated with or without addition of 2i for 2 days were subjected to qRT-PCR analysis.
