SHIP2 phosphatase activity and capacity to interact with Mena are required for cell invasion in vivo. (A) Mice injected (tail vein) with a MDA-MB-231-1833-TR bone metastatic variant expressing vector (pLKO.1), SHIP2 shRNA, or SHIP2 shRNA plus GFP-SHIP2-WT, GFP-SHIP2-D608A, or GFP-SHIP2ΔFP4 were imaged to detect luciferase activity. Bioluminescence images of two representative mice per condition acquired immediately and 1 and 7 wk after injection are depicted. (B) Tumor growth within lungs, for mice injected with cells as indicated in A, was quantified and expressed as fold change of initial inoculum on day 0. (C) SDS-PAGE followed by Western blot analysis depicts protein levels of SHIP2 between MDA-MB-231-1833-TR cells transduced with pLKO.1 and shSHIP2 as well as exogenous rescue levels of GFP-SHIP2-WT, GFP-SHIP2-D608A, and GFP-SHIP2ΔFP4. (D) Number of lung lesions per mouse was quantified based on hematoxylin and eosin staining of serial step sections of paraffin embedded tissue. (E) Table summarizes the cells injected via tail vein and their respective take rate for tumor growth at the lungs. (F) Table summarizes the cells injected via tail vein and their respective tumor spread and growth rate outside lungs. (G) Representative hematoxylin and eosin staining of a single section of all five lobes of mouse lung shows characteristic lesions among different conditions. All quantified data indicate the mean values ± SE. *, P < 0.05; NS, not statistically significant. Bars, 4 mm.