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. Author manuscript; available in PMC: 2008 Feb 3.
Published in final edited form as: Biochem Biophys Res Commun. 2006 Dec 11;353(1):189–194. doi: 10.1016/j.bbrc.2006.12.018

Fig. 1.

Fig. 1

Fig. 1

Fig. 1

Enhancement of GAS6 mRNA expression in normal mammary epithelial cells and ER positive breast cancer cells by estrogen. a). Induction of GAS6 by estrogen in mammary glands. Ovariectomized mice were treated with 17β-estradiol (E2) for 9 hours or 24 hours and mammary glands were then isolated for the preparation of total RNA. Total RNA (10 μg) was hybridized with radiolabeled anti-sense GAS6 probe generated by in vitro transcription. Glyceraldehyde-3-phosphate dehydrogenase probe was included in each hybridization reaction as control. Nuclear resistant fragments were resolved on polyacrylamide-urea gels. b). GAS6 is mainly expressed in mammary epithelial cells. RT-PCR was performed with total RNA prepared from the mammary epithelial cells and stromal cells, which had been separated by collagenase digestion and Percoll gradient centrifugation. c). GAS6 is induced by 17β-estradiol (E2) in MCF-7 cells. Total RNA was made form MCF-7 cell treated with E2 or control vehicle for 3 hours. The GAS6 and pS2 mRNA levels were quantified by SYBR Green real-time PCR and normalized to 18S ribosomal RNA.