Ectopic expression of SHIP-2 in SHIP-1^-/-BMMs. (A) BMMs from SHIP-1^+/+ and SHIP-1^-/- animals were tested for the expression of SHIP-1 and SHIP-2 by Western blotting. (B) Using the Nucleofector, BMMs were transfected with plasmids encoding GFP (left: immunofluorescence image; right: phase contrast image of the same field). Images were obtained using an inverted fluorescence microscope with ×40 magnification. (C) SHIP-1 knockout BMMs were transiently transfected with empty vector alone, wild-type SHIP-2 (SHIP-2 WT), or catalytically deficient SHIP-2 (SHIP-2 D608A). GFP was cotransfected as a marker for transfection. Whole cell lysates from the transfectants were separated on 10% SDS-PAGE and transferred to nitrocellulose membrane, which was blotted with anti-SHIP-2 or anti-GFP antibody. These results are representative of 3 independent experiments.