Simultaneous detection of the alternatively spliced mRNA species of Ang-1 in CHRF cells by RNase protection assay.

Schematic diagrams of the strategy for determining the alternatively spliced mRNA of Ang-1 are shown on the upper panels of A and B. The thick lines represent fragments derived from various forms of Ang-1 that could be detected by this assay. The alternatively spliced parts of Ang-1 are shown as thin lines. (A) The designed 262-bp antisense probe is derived from the 1.3-kb isoform. Lane 1 shows the RNase-protected 262-bp product, indicating the presence of the 1.3-kb isoform and the 219-bp product representing the 1.5-kb and 0.9-kb isoforms. The 43-bp fragment (not shown) is small and seen only after a longer time exposure. Lane 2 shows the RNA extracted from DU145 cells as a negative control. (B) The designed 249-bp antisense probe is derived from the 0.9-kb isoform. The detected 249-bp band indicates the presence of the 0.9-kb isoform. The 146-bp and 103-bp bands represent the 1.5-kb or 1.3-kb isoforms.