Figure 8.

Radil is required for breast cancer cell metastasis. (A) MDA-MB-231 cells stably expressing Renilla luciferase were transduced with scrambled shRNA or Radil shRNA #4 and their migratory potential was tested using Transwell assays before injection in mice. (B) MDA-MB-231 cells were injected via tail vein in NOD-SCID mice. Metastasis and homing of cells to the lungs was monitored over time using in vivo bioluminescence imaging. Shown are images from day 35. (C) Mice were sacrificed 45 d after inoculations. Lungs from mice injected with saline or with scrambled shRNA or Radil shRNA–expressing cells were collected, fixed, and stained with Bouin’s solution. Shown are representative images from each condition (top). Quantification of the visible tumor nodules is provided in the table (below). Scrambled shRNA, n = 12; Radil shRNA #4, n = 11; statistics, Student’s t test. (D) Total number of nodules counted on the lungs of each animal. Error bars, ±SEM. (E) In vitro proliferation of MDA-MB-231 cells treated with scrambled or Radil shRNA #4. (F) Quantification of tumor end volume (in cm³) on day 31. Scrambled shRNA, n = 5; Radil shRNA #4, n = 5. Red line denotes median value. Bars, interquartile range. (G) Quantification of tumor end weight (in grams) on day 31.