CEA_SGF:E00004#TetOFF_Stat3F

The goal of this study was to identify the target genes of Stat3 involved in murine ES cell self-renewal. To achieve this goal, we used the Gs2 ES cell line expressing an inducible dominant negative mutant of Stat3 (Stat3F) in wich Y705 is mutated to phenilalanine. The Gs2 ES cells are routinely maintained in the presence of LIF 1000 U/ml and tetracycline 1µg/ml (LIF/ON). In this condition, Stat3F is not expressed and the cells are maintained in an undifferentiated phenotype. Upon tetracycline removal but inthe presence of LIF (Tet OFF condition) Stat3F is expressed and the cells differentiate despite the presence of LIF. We thus performed microarray analysis of the transcriptome of ES cells after 16h, 24h and 48h after Tetracycline removal (OFF16; OFF24 and OFF48) respectively)and compared them to Gs2 ES cells maintained in the continuous presence of LIF and tetracyclin (LIF/ON). Keywords: time-course