WNT5A Inhibits Metastasis and Alters Splicing of Cd44 in Breast Cancer Cells
Figure 5
(A) Genomic structure of mouse Cd44 is shown. Cd44 has 9 non-variable exons numbered 1 to 9 (large). After non-variable exon 5 there are 10 variable exons numbered v1 to v10 (small). Primers used for RT-PCR are shown as small arrows on top of the genome structure. All PCR reactions used the same reverse primer in exon 6. Cd44s, the non-variable form of Cdd4, is the main PCR product amplified using the Cd44s primer in exon 5. Variable exons are amplified using primers in exon v4, v6 and v9. (B) RT-PCR for various Cd44 transcripts is shown (Cd44s, v4, v6, and v9). Expression of WNT5A was confirmed using primers to human WNT5A. Gapdh was used as a normalization control. (C) PCR was carried out for varying cycles to determine the linear range of product formation. Amplification of Cd44s is shown. Gapdh is used as a control for normalization. (D) Pixel density of bands from the images of the stained gels was determined and normalized to that of Gapdh. Bands in the linear range of product formation were used. Three separate experiments were analyzed. The vector was set to 1.0 and the relative levels of Cd44s, V4, V6 and V9 were determined. The average and standard deviation are shown on the graph. * = T-test p-value <0.05.