Figure 7.

Radil and KIF14 are required for MDA-MB-231 cell migration and invasion. (A) MDA-MB-231 cells were transduced with the indicated shRNAs. 72 h after transduction 5 × 10⁴ cells were seeded on the upper chamber of Transwells and 20% fetal bovine serum (FBS) in DMEM was applied to the lower chamber. Cells were allowed to migrate for 10–12 h and migrated cells counted from pictures of 4–5 random fields. (B) Cell invasion was assessed as above except Transwells coated with 1 µg/ml of matrigel were used. Cells were allowed to invade through the matrigel for 20 h. (C) MDA-MB-231 cells stably expressing the shRNA-resistant murine full-length mRadil or mRadilΔPDZ were transduced with scrambled, or two different Radil shRNAs. The migratory potential of these cells was assessed as described above. Expression levels of FLAG-mRadil and FLAG-mRadilΔPDZ are shown on the right. Cortactin used as loading control. (D) Transwell cell migration and (E) invasion assays with cells expressing KIF14 shRNAs. Cells were processed as above. (F) MDA-MB-231 cells were transduced with scrambled shRNA or KIF14 shRNA #816 in the presence of FLAG-GFP, eGFP-KIF14, or eGFP-KIF14-IQAA. The different cells were then subjected to Transwell assays. Expression levels of eGFP-KIF14 and eGFP-KIF14-IQAA are shown on the right. KIF14 shRNA #816 targets the 3′ UTR and was thus used for rescue experiments. Bars, 50 µm. Error bars, mean ± SEM.