Effects of AKT, GSK3 and MEK inhibitors on the expression of endogenous Sox2.

<p>1.5×10⁶ i-OSKM-ESCs were cultured in 100 mm dishes with or without 4 μg/ml Dox. (A) 24 hours after the cells were plated in the absence or presence of Dox, 5 µM AKTiV and 3 µM CHIR were added to the cells for an additional 24 hours where indicated. (B) 24 hours after the cells were plated in the absence or presence of Dox, 5 µM AKTiV and 0.4 µM MEKi were added to the cells for an additional 24 hours where indicated. For A-B, nuclear and cytoplasmic protein extracts were prepared from the cells and equal amounts of nuclear and cytoplasmic protein were loaded into each well of an SDS-PAGE. Western blot analysis was performed by sequentially probing for Sox2 and HDAC1 in A and B. In each case, HDAC1 served as a protein loading control.