Quantitative analysis of reticulocyte 4.1R messenger RNA (mRNA).
(A) RNase protection assay. In the top panel is an autoradiograph showing phosphorus 32 (32P)-labeled protected fragments. Mk indicates size marker (32P end-labeled pBR 322 MspI fragments); P, undigested probe; Ct1 and Ct2, control mRNAs; and Y, yeast RNA used as negative control. The middle panel shows a schematic representation of the probe and the expected protected fragments. The bottom panel shows the 4.1R mRNA quantitation measured as a normalized ratio of total 4.1 to band 3. (B) Quantitative RT-PCR using protein 4.2 mRNA as the internal control. Three time points (20, 22, and 24 cycles) were considered. Mk indicates 100-base-pair size marker. Total 4.1R mRNA amounts are presented as ratios of 4.1R to 4.2.
