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. 2015 Sep 22;6(33):34691–34703. doi: 10.18632/oncotarget.5787

Figure 2. H19 knockdown decreases pluripotency of hES and hEC and promotes early differentiation.

Figure 2

A.-E. Down-regulation of H19 expression decreases the RNA and protein expression levels of Oct4 and Nanog in hES and hEC cell lines. A. qPCR of RNA from NCCIT cells for Oct4 (n = 10) and Nanog (n = 9). B. qPCR of RNA from NT2 cells for Oct4 (n = 3) and Nanog (n = 3). C. qPCR of RNA from HES-1 cells for Oct4 (n = 2) and Nanog (n = 2). D. Western blot of Protein lysates from NCCIT cells for Oct4 and Nanog E. Western blot of Protein lysates from NT2 cells for Oct4 and Nanog F. H19 knockdown in NCCIT cells reduces the expression levels of surface markers TRA-1-60 and TRA-1-81, as analyzed by FACS (n = 2).G. H19 down-regulation induces an increase of SSEA1 expression in shH19 transduced NCCIT cells, as analyzed by FACS (n = 4).H. H19 down-regulation in transduced NCCIT cells induces a decrease of Oct4 (green) and an increase of Nestin (Red) expression, as assessed by immunofluorescence (Nuclear counterstaining with Dapi (Blue)). Data are represented as mean ± SD; *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001.