We next used nucleofection to analyze the function of Sox2 in maintaining hES cell pluripotency. Reduction of Sox2 expression by RNA interference resulted in a loss of the hES cell state as demonstrated by altered morphology, loss of stem cell antigen expression, and differentiation into trophectoderm. Downregulation of Sox2 also decreased Oct4/Nanog expression, suggesting a coordinated role for these factors in regulating pluripotency. These results demonstrate that Sox2 is required to preserve the hES cell state.Alexa fluor 488 rabbit anti-goat IgG (Invitrogen, 1 :400 dilution) was added for 1 hour at room temperature. ... BrdU-loaded for 2 hours at 37AdC, then fixed, blocked , and DNase treated according the manufacturera#39;s instructions (BD Biosciences).
|Title||:||Dissecting Cell Fate Decisions in Human Embryonic Stem Cells|
|Author||:||Kristi A. Hohenstein|
|Publisher||:||ProQuest - 2009|