Figure 6
Ret contributes to motility and to a lesser extent, colony formation by the LM7 cell line. (a–e) The first validation strategy compared the effects of the siRNA pool with that of the individual duplexes. Knockdown efficiency was assessed by real-time PCR measurements of Ret mRNA levels (mean±s.e.m. of three PCR well replicates) (a). Effects of the siRNAs on in vitro phenotypes were assessed by measuring motility (b), colony formation (c), invasion (d) and cell growth (e). Results are presented as mean±s.e.m. of the number of replicates indicated in the Materials and methods for each assay. (f–h) The second validation strategy compared the effects of an antisense LNA/DNA gapmer targeting Ret with that of the siRNA pool. Knockdown efficiency by the antisense or siRNA pool was assessed by measuring Ret mRNA levels by real-time PCR, comparing levels with cells treated with either the non-targeting antisense LNA/DNA gapmer or the siRNA non-targeting pool (f). Effects of the antisense LNA/DNA gapmer and the siRNA pool on in vitro phenotypes were assessed by motility (g) and colony formation (h). Results are presented as mean±s.e.m. of four independent experiments.
