Activin-A stimulation of ES cells was performed as follows. ES cell lines were started on gelatinized plates in ESC Growth Media. Cultures were then grown for 2 days in ESC Growth Media without LIF. On the third day, cells were cultured in low serum ESC Growth Media without LIF (same as ESC Growth Media except 0.1% FCS is used instead of 15% FCS) in the presence or absence of 30 ng/ml activin-A (R & D Systems) overnight. Cells were processed using TriReagent according to manufactures standard protocol.
