QRT-PCR analysis of total RNA from human primary mammary epithelial cells (AG1132B) and from established breast cancer cell lines, Hs578T. MDA-MB231, and T47D cells. 1 μg of total RNA was used to perform reverse transcription reactions (RTs) and 1 ul of the RT reaction was used to set up qRT-PCR reactions in triplicates using RASSF1A and RASSF1C specific primers. All qRT-PCR reactions were set in triplicates and Cyclophillin was used as an internal loading control and used to normalize the relative expression levels using the 2-ΔΔ method . The RT-PCR analysis shows that RASSF1A expression was down and RASSF1C expression is up in established breast cancer cell lines compared the primary mammary epithelial cells.