Figure 2. Pimozide reduces tumor burden, cell proliferation, and the number of lung metastases in a nude mice xenograft model system.

(A) In vivo bioluminescence imaging system (IVIS) of MDA-MB-231-Luc (D3H2LN) xenografts in SCID mice. Ventral (upper left panels) images taken over time from representative mice (10 imaged). Upper right panels show the Region of Interest (ROI) in localized tumors from representative mice (10 imaged). Pseudo color scale bars were consistent for all imaged ventral views in order to show relative changes at metastatic sites over time, lower left panel images show metastatic foci in control mice (G2NT), and the lower right panel shows the ROI in different mice (red arrow). (B) Characterization of tumors in untreated (PBS) and treated (Pimozide) groups by plotting tumor volume (mm³), tumor occurrence (%), ROI (photons/sec) and lung metastatic foci numbers with significance values at autopsy. Data shown are means ± SD (Student’s t test ^*P <0.05, ^**P < 0.01). (C) Hematoxylin & eosin (H&E) staining of xenograft tumors in untreated (NT) (a), earlier treated (TE) (b) and late treated (TL) (c) mice. Magnification 200x. Scale bar 100 μm. Immunohistochemical staining for Ki67 (d,e,f). Magnification 200x. Scale bar 100 μm, cleaved caspase-3 (g,h,i). Magnification 200x. Scale bar 100 μm (g). Magnification 400x. Scale bar 50 μm (h, i) and Ran (j,k,l) in NT, TE and TL mice. Magnification 200x.Scale bar 100 μm (D) H&E staining of representative lung metastases from WT, NT, and TE mice. Ran immunostaining was assessed in wild type lung and lung metastases (NT and TE), magnification 200x. Scale bar 100 μm. (E & F) Immunohistochemical staining for AKT in MDA-MB-231 xenograft tumors. (E) Immunohistochemical staining for AKT (Pan). Scale bar 100 μm. (F) AKT-isoforms (AKT1, AKT2) in untreated and treated mice breast xenografts tumors showing a representative result for 10 sections per group. Magnification 100x. Scale bar 100 μm.