FIGURE 6.

C/EBPβ mRNA half-life of βwt and βd/s cell lines. At 24 h after induction of differentiation, doxycycline was added to the culture medium, and total RNA was isolated with respect to time. Real-time PCR analysis was utilized to quantify the C/EBP βwt (Bw/t) and βd/s (Bd/s) mRNAs remaining after addition of the doxycycline. The half-lives of the C/EBPβ mRNAs were determined graphically using a plot of log[mRNA] versus time as described under “Experimental Procedures.” A, results of the real-time analysis. Results are plotted as the mean ± S.D. of two independent experiments analyzed in triplicate. B, half-life plot of real-time results. βwt (○) and βd/s (▵) mRNAs remaining after addition of doxycycline with respect to time are shown. The equation y = ae^-bx was fitted to the data, and half-lives were calculated. The experiment was performed twice with similar results.