Comparison of the Invader RNA assay, RNase protection assay, and RT-PCR assay in the quantification of FGFR2 transcripts. pI12DE:Wt, which is described in Figure 4A, was transfected into DT3 cells. Total RNA was extracted and analyzed with the Invader RNA assay, RNase protection assay, or RT-PCR to quantify U-IIIb-IIIc-D RNA (gray bars), U-IIIb-D RNA (white bars), or U-IIIc-D RNA (black bars). Error bars represent one standard deviation. The percent inclusion of a product (e.g., U-IIIb-D) among transcripts was calculated as follows: percent product (e.g., U-IIIb-D) inclusion = {[product (e.g., U-IIIb-D)]/([U-IIIb-D)] + [U-IIIc-D] + [U-IIIb-IIIc-D])} × 100. This RPA cannot accurately quantify the skipped product (U-D) because the probe products protected by this splice variant were also generated from the protection of other spliced products (i.e., U-IIIb-D and U-IIIc-D).
