PCR Base Line Subtracted CF RFU

1. A B PCR Base Line Subtracted CF RFU PCR Base Line Subtracted CF RFU Cycle Cycle Supplementary Figure S2: Gene expression of dopamine transporter (DAT) and norepinephrine Transporter (NET) in rat pheochromocytoma cells (PC12) and human embryonic kidney 293 cells (HEK) using real time PCR Real time PCR was preformed according to the protocol of QuantiFast 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) PC12 , HEK and the PC12 (old generation ATCC from Israel) . Violet= NET expression of HEK; Green= NET expression of ECACC PC12; light blue= NET expression of Israel PC12; dark blue = DAT expression of ECACC PC12. All other lines under baseline = DAT expression of Israel PC12 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 mRNA and therefore can be used as a positive control. The undifferentiated PC12 cells obtained from Israel express no DAT but show a high NET mRNA expression. The undifferentiated ECACC PC12 cells express DAT and show also a high NET mRNA expression. Annotation: CF RFU, Relative Fluorescent Units.