Time-course analysis was performed by using shRNA directed against Brg1 . (A) Transfection of shRNA against Brg1 showed up to 90% downregulation of Brg1 in ESD3 cells compared with embryonic stem (ES) cells transfected with control shRNA. (B) Transfection of sh Brg1 led to an increase in Oct4 levels at 24 h posttransfection but a decrease in Oct4 levels at 48 and 72 h posttransfection with sh Brg1 , compared with cells transfected with control shRNA. (C) Nanog levels remained elevated for up to 72 h after knockdown of BRG1. (D) siRNA-mediated BRG1 showed 85% knockdown efficiency. (E) Western blot analysis was carried out at 48 h posttransfection using cell lysates prepared from mouse ES cells. BRG1 protein levels were detected by using polyclonal anti-BRG1 (N15) antibody; β-TUBULIN was used as a loading control. (F) Levels of Oct4 and Nanog were increased and Sox2 levels were decreased in ES cells at 48 h posttransfection.