Figure 1

Distinctive quantification of ccn1 and ccn2 mRNA by real-time RT-PCR. A. Primers used for real-time PCR and the structures of human ccn1 and ccn2 mRNAs. Schematic representations are illustrated in reference to the modular structure of human CCN1 and CCN2 (stippled boxes). The small open circle and AAAAA at the left and right ends denote the 5'-cap structure and poly-A tail, respectively. Names, locations for recognition, nucleotide sequence of the primers, and the expected sizes of the PCR products are given. Abbreviations: IGFBP, insulin-like growth factor binding module; VWC, von Willebrand factor type C module; TSP1, thrombospondin type 1 repeat; CT, C-terminal module. B. The CCN1 (219 bp), CCN2 (153 bp), and β-actin (101 bp) PCR products amplified by LightCycler were analyzed by agarose electrophoresis. C. Melting curve analysis of the RT- PCR products of ccn1 and ccn2. Melting curves were acquired after 45 cycles of amplification. Melting curve pattern is displayed on the left panel, where fluorescent intensity (F1) from SYBR green is plotted against temperature. Melting peak pattern can be found on the right panel, in which the decrement of F1 is plotted against temperature.