FIG. 6.

Total Src and activated Src and PYK2 are mislocalized in PLCγ2^−/− osteoclasts. WT BMDM, retrovirally transduced with pMX empty vector (A, D, G, J, and M), and PLCγ2^−/− BMDM, retrovirally transduced with pMX-NFAT2 (B, E, H, K, and N) or pMX-PLCγ2 (C, F, I, L, and O), were grown in differentiation medium for 5 to 7 days to allow osteoclast differentiation. Cells were then fixed and stained with phalloidin (A to C) or incubated with antibodies to PYK2 (D to F), p402 PYK2 (G to I), Src (J to L), or p416 Src (M to O). An asterisk marks each mature osteoclast present in a field. All other cells are immature preOCs, macrophages, or stromal cells. Arrows mark the area at or near the actin sealing ring in mature osteoclasts. In panels H and N, a dotted line delineates the edge of the mature osteoclast. Images were captured using an Olympus IX70 inverted microscope (60× APO, 1.4-numerical-aperture lens), with a CoolSnap camera, and analyzed using Metamorph software. Scale bar, 5 μm.