FIG. 8.

Translation antitermination mutations reduce the stability of hβ-globin mRNA in intact erythroid cells. (A) Representative two-probe RPA. RNA prepared from bone marrow progenitors (B) and peripheral blood reticulocytes (P) obtained from representative mice containing the β^WT transgene (lanes 3 and 4) or the β^RT transgene (lanes 5 and 6) was analyzed using ³²P-labeled probes complementary to hβ-globin (hβ) and mα-globin (mα) mRNAs. The migration of the protected mα and hβ probe fragments is indicated by control reactions utilizing either of the probes alone (lanes 1 and 2). The composition of each reaction and the migration of the protected fragments are shown above and to the left of the autoradiograph, respectively. Control lanes indicating functional probe excess, performed for every experiment, were cropped from the figure to maintain clarity. (B) Stabilities of β^WT and β^RT mRNAs in multiple transgenic lines. The average stabilities of β^WT and β^RT mRNAs in each of five and one independent line, respectively, are plotted (●). For each line, a minimum of two mice were studied on at least two separate occasions using the two-probe (hβ and mα) RPA. The stability of each mRNA, averaged across all lines, is indicated as a bar; the stability of mα-globin mRNA, defined as 1.0, is denoted by a dashed line.