PCR Base Line Subtracted CF RFU A Cycle

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PCR Base Line Subtracted CF RFU A Cycle B Cycle Supplementary Figure S 2: Gene expression of dopamine transporter (DAT) and norepinephrine Transporter (NET) in rat pheochromocytoma cells (PC 12) and human embryonic kidney 293 cells (HEK) using real time PCR Real time PCR was preformed according to the protocol of Quanti. Fast SYBR Green PCR Kit (# 204056). All cells were undifferentiated cells (no nerve growth factor was used). A. Representative gene expression of DAT and NET in the European Collection of Cell Cultures (ECACC) PC 12 , HEK and the PC 12 (old generation ATCC from Israel). Violet= NET expression of HEK; Green= NET expression of ECACC PC 12; light blue= NET expression of Israel PC 12; dark blue = DAT expression of ECACC PC 12. All other lines under baseline = DAT expression of Israel PC 12 and negative controls. B. Representative DAT expression of HEK cells as a positive control (green and red- duplicate). All other lines under baseline= negative controls. Using real time RT-PCR for detection of DAT and NET gene expression, we could show that HEK cells expresses DAT m. RNA and therefore can be used as a positive control. The undifferentiated PC 12 cells obtained from Israel express no DAT but show a high NET m. RNA expression. The undifferentiated ECACC PC 12 cells express DAT and show also a high NET m. RNA expression. Annotation: CF RFU, Relative Fluorescent Units.

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