Figure 6

Small ubiquitin-related modifier (SUMO)-interacting motif (SIM) of Oct4 is crucial for the interaction with ERG-associated protein with SET domain (ESET) and the repression of Cdx2. (a) Figure depicts SIM in Oct4 (top panel) and the mutations (red alphabet) in the Oct4-lacking SIM (-ΔSIM) (bottom panel). (b) Expression vectors indicated were transfected in 293T cells and immunoprecipitant (IP) from anti-Flag antibody and input were subjected to western blot (WB) with anti-haemagglutinin (HA) (ESET, top panel) and anti-Flag (Oct4 or Oct4-ΔSIM, bottom panel) antibodies. (c) Quantitative polymerase chain reaction (Q-PCR) analysis of Cdx2 (top), Oct4 (middle) and Eset (bottom) in ZHBTc4 ES cells that were either non-treated or treated with tetracycline (Tc) to deplete the endogenous Oct4 and transfected with the indicated plasmids, relative to control embryonic stem (ES) cells which was set as 1.0 (lane 1) after normalising against Gapdh. An empty vector was used as a transfection control in lane 2. Error bars represent standard deviation (SD) of the average and median of four different fractions of cells of different green fluorescent protein (GFP) intensity except for control ES cells (lane 1) where error bar represents the SD of three technical replicates.