RPA demonstrates up-regulated VCAM/GAPDH mRNA ratios in the lungs of transgenic NY-S and NY-S/S^Antilles mice and LPS-treated control mice (18 hours after LPS injection) compared with normal lung controls.

Lung homogenates were prepared from 3 mice in each group. Total RNA was extracted from the lungs of sickle and normal mice. RNA protection assays (RPA) were performed with 10 μg extracted mouse lung RNA using VCAM and GAPDH antisense RNA probe templates labeled with [³²P]UTP. The ³²P-labeled antisense VCAM and GAPDH RNA probes were hybridized to lung mRNA overnight and then digested with RNAase. Protected RNA fragments were separated by electrophoresis. Radioactive bands corresponding to protected VCAM and GAPDH mRNA were quantified by phosphorescence densitometry. The figure shows the bands from one representative lung from each group and a summary bar graph. The bar graph plots the mean ± SD VCAM/GAPDH mRNA ratios for each mouse model expressed as a percentage of the VCAM/GAPDH mRNA ratio in normal control mice (n = 3). bp indicates base pair. **P ≤ .01; ***P ≤ .001.