Differential regulation of the human embryonic targets of NANOG, OCT4, and SOX2 (NOS targets) in OCT4-transduced breast cells (OTBCs). (a) Expression of selected NOS targets across the parental cell line p86, the OTBCs86-L1 cell line, and the same line OTBCs86-L1 transfected with a short interfering RNA specific for OCT4. Efficient knockdown of OCT4 was validated by Western blot. Each colored square on the left panels represents the mean relative transcript abundance (in log2 space); highest expression is red, average expression is black, and lowest expression is green. Data represent the average of three independent experiments. (b) Detection of self-renewal transcription factors OCT4, NANOG, SOX2, and ZIC1 expression by quantitative real-time polymerase chain reaction (qRT-PCR). Control cell lines used for gene expression analyses include human dermal fibroblasts and human embryonic stem cells (hESCs). The mRNA levels were normalized to those of the parental cell lines (p86 and p48). Bar graph represents the mean ± standard deviation (SD) of three independent experiments. (c) Detection of tumor suppressor gene expression by qRT-PCR. Genes analyzed include p21WAF1/cip1, Dickkopf-related protein 1 (DKK1), methylguanine-DNA methyltransferase (MGMT), Maspin (SERPINB5), and E-cadherin (CDH1). The mRNA levels were normalized to those of the parental cell lines (p86 and p48). Bar graph represents the mean ± SD of three independent experiments. NOS, NANOG, OCT4, and SOX2 targets.
Beltran et al. Breast Cancer Research 2011 13:R94 doi:10.1186/bcr3019