Research Paper Volume 2, Issue 11 pp 815—822
Core transcription factors, Oct4, Sox2 and Nanog, individually form complexes with nucleophosmin (Npm1) to control embryonic stem (ES) cell fate determination
Figure 5. Tpt1 and Npm1 are involved in ES cell maintenance. Involvement of Tpt1 and Npm1 in ES cell maintenance was investigated using shRNA constructs against each gene and analyzed by qPCR 48 h post transfection. Downregulation of Tpt1 (blue bars) resulted in a minor increase in Oct4 and Brachyury (Bra) levels, a decrease in GATA4 levels, whereas levels of Sox2 and Nestin (Nes) increased significantly, indicative of involvement in ectodermal differentiation. Downregulation of Npm1 (black bars) resulted in minor increases in Oct4 and Sox2 levels, decreased GATA4 levels, while increased Brachyury (Bra) levels, indicative of involvement in mesodermal differentiation. All experiments were done in triplicates and normalized first to GAPDH (reference gene), and then to corresponding negative control shRNA constructs.