Fig. 2.
Effect of treatment with acidic medium on the half-life of the endogenous PEPCK mRNA in LLC-PK1-F+-9C cells. Confluent cultures of cells were treated for 12 h in either normal, pH 7.4 (•——•) or acidic, pH 6.9 (□——□) medium. RNA was isolated at various times after addition of 8 μg/ml actinomycin D. The relative levels of PEPCK and GAPDH mRNAs were quantified by real-time RT-PCR. The level of PEPCK mRNA was divided by the corresponding level of GAPDH mRNA. The log of the normalized data was then plotted against the time after the addition of actinomycin D. The reported data are means ± SE of duplicate experiments each of which contained triplicate samples for each time point.