FIGURE 1.

Ash2l is critical for maintaining ES cell pluripotency. a, mouse ES cells were treated with retinoic acid (RA) and then harvested at the indicated time points for Western blotting with the indicated antibodies. Tubulin served as loading control. b, mouse ES cells were transfected with control (ctrl) or two different Ash2l siRNA oligonucleotides for knockdown. Extracts were collected after 4 days for Western blotting with the indicated antibodies. GAPDH served as loading control. Relative levels of Ash2l after normalization with loading controls were determined using NIH ImageJ software. c–e, self-renewal was impaired in Ash2l knockdown cells. Ash2l RNAi treated cells from (b) were examined for expression of the indicated self-renewal genes by RT-qPCR (c), alkaline phosphatase activities (d), and expression of the indicated lineage markers by RT-qPCR (e). Error bars indicate S.E. (n = 3).