Fig. 4. Decreased expression of human MIRTD in rat PC12 cells carrying the human DLST gene with haplotype ac. PC12 cells were cotransfected with the cosmid containing the ac- or gc-type DLST gene and the geneticin-resistance gene using Lipofectin (Invitrogen). Transfectants were enriched by selecting geneticin-resistant cells for 14 days and then poly(A)⁺ RNA was prepared from the transfectant pools. Amounts of human MIRTD and DLST mRNAs were measured by real-time quantitative RT–PCR. (A) Profiles of the real-time quantitative RT–PCR showing increases of PCR products derived from human DLST and MIRTD mRNAs. (B) Profiles of the real-time quantitative RT–PCR using poly(A)⁺ RNA from untransfected PC12 cells, detecting neither DLST nor MIRTD mRNA. (C) Relative values of MIRTD to DLST mRNA are shown with the mean ± SD obtained from four independent experiments. The difference was significant for 1 versus 3 (p = 0.039), 1 versus 4 (p = 0.049), 2 versus 3 (p = 0.016) and 2 versus 4 (p = 0.016).