Fig. 3

Tissue distribution of normal hCNT1-WT and intron retention variant transcript hCNT1-IR. For quantitative measurement of mRNA, DNase-treated human tissue total RNA human tissue total RNA panel was purchased from ClonTech. cDNA was synthesized using SuperScript II Reverse Transcriptase (Invitrogen). The PCR reactions using cDNA were performed in a LightCycler system (Roche) under the following conditions: 95 °C for 5 min, followed by 40 cycles at 95 °C for 15 s, 60 °C for 30 s, and 72 °C for 10 s. The relative level of amplified mRNA was normalized to mRNA expression of the housekeeping gene GAPDH (glyceraldehyde 3-phosphate dehydrogenase). Gene expression was evaluated by the ΔCT method relative to the reference gene. All samples were amplified in duplicate and two non-template controls per primer pair were included in each run. a qRT-PCR analysis of different human tissue RNA that using primers for normal hCNT1 transcript. b qRT-PCR analysis of different human tissue RNA using primers for intron retention hCNT1-IR splice variant transcript. Data are the mean ± SEM for three experiments run in duplicates