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We chose NTera2 cells, a pluripotent human embryonal carcinoma cell line derived from an adult testicular GC cancer patient because they express markers of fetal GC, such as TFAP2C and hence they are commonly used as a fetal GC model (Hoei-Hansen et al.
AB5535) and GC subpopulations, namely gonocytes (TFAP2C; Santa Cruz Biotechology; catalog no.
Fetal testis sections (one per cultured or xenografted tissue piece) were co-immunostained for sex-determining region Y-box 9 (SOX9; Sertoli cells), transcription-factor activating enhancer-binding protein 2 (TFAP2C; gonocytes), and melanoma-associated antigen 4 (MAGEA4; prespermatogonia).
For fetal ovary analyses, sections (one per cultured piece) were co-immunostained for TFAP2C (to identify all GC) and Hoechst as a counterstain.
Expression of epigenetic regulators in human (TET1, EZH2, DNMT3a, DNMT3b) and rat (Tetl, Ezh2, Dnmt3a and Dnmt3b), in addition to germ cell pluripotency (POU5F1, TFAP2C and NANOG) genes were determined using the Roche Universal Probe Library (Roche Applied Sciences) (see "NTERA2--primer sequences for gene expression analysis" in the Supplemental Material for primer sequences).
We therefore investigated whether expression of the established GC pluripotency markers POU5F1, TFAP2C, and NANOG (Rajpert-De Meyts 2006) differed from vehicle controls in NTera2 cells cultured with acetaminophen, ibuprofen, or EP2 + EP4 antagonists (10 [micro]M each).
In the first case, we have for instance NFYA, NFYB, and NFYC; whereas in the latter case, we have TFAP2A, TFAP2C, and MAZ.
Those genes are NFYB, NFYC, and NFYA for activators and MAZ, TFAP2C, and TFAP2A for repressors.
Gu et al., "EGFR is regulated by TFAP2C in luminal breast cancer and is a target for vandetanib," Molecular Cancer Therapeutics, vol.
They focused on three key transcription factors that they found interact physically with each other--Elf5, Eomes and Tfap2c.
Too much of Elf5 or Tfap2c in proportion to Eomes triggered TSCs to differentiate.
They identified three genes (Gata3, Eomes and Tfap2c) that when introduced into skin cells, they could initiate a cell fate program that completely converted the skin cells into stable and fully functional placenta-generating cells.
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