Fig. 1

Rictor ablation decreases spontaneous and experimental metastasis in transgenic mammary tumors. a H&E staining of lungs from tumor-bearing MMTV-NIC mice. Arrows indicate metastases. Representative images are shown, original magnification, ×40. b Left panel: percentage of tumor-bearing mice harboring lung metastases. Non-tumor bearing mice were excluded from this analysis. Error bars represent the average of two experimental repeats. The first trial sample numbers were 7 Rictor ^+/+ NIC and 5 Rictor ^FL/FL NIC. The second trial numbers were 5 Rictor ^+/+ NIC and 4 Rictor ^FL/FL NIC. Right panel: number of lung metastases per tumor-bearing mouse was determined in H&E-stained sections through the lung. Midlines are the average number (± S.D.) of metastases/mouse. Each data point represents one tumor-bearing mouse; N = 12 Rictor ^+/+ NIC and 9 Rictor ^FL/FL NIC. c-d Dissociated Rictor ^+/+ NIC and Rictor ^FL/FL NIC tumors (N = 3) were partitioned for assessment by western analysis (c) or for venous delivery to wild-type (WT) FVB mice by (two recipient mice per donor). At 8 weeks, lungs were harvested from each recipient and assessed for metastatic lesions by whole-mount hematoxylin staining (d). Midlines are the average number (± S.D.) of metastases/mouse. Each data point represents one mouse. e-g MMTV-Neu-derived mouse mammary tumor cells retrovirally transduced with pBABE or pBABE-Rictor. e Cells were treated for 4 h with lapatinib, then assessed by western analysis using the antibodies indicated (left). f Venous delivery of cells to WT FVB recipient mice was followed 8 weeks later with examination of lungs from each recipient for metastatic lesions. Representative images are shown, original magnification × 40. Upper panels: histological sections. Lower panels: Rictor immunohistochemical analysis (IHC). g Left panel: percentage of mice harboring lung metastases, one trial, N = 12 per group. Right panel: number of lung metastases/mouse was determined as in b. HER2 human epidermal growth factor receptor 2