Fig. 6. Localization of Dd-EF2H in ef-2^AS, ef2-null and parental Ax-2 cells. (A-H) Cells were harvested at the exponential growth phase, fixed in absolute methanol and double-stained with the anti-Dd-EF2H antibody or non-immune serum, and then with DAPI, as noted in Materials and Methods. In Ax-2 cells (A-C), the cytoplasm was immuno-stained by the anti-Dd-EF2H antibody (B). Surprisingly, in ef-2^AS cells (D-F), immunostaining of cytoplasmic granules is retained. Higher magnification of ef2-null cells double-stained with the anti-Dd-EF2H antibody and DAPI (G,H) indicates that the distribution of cytopasmic granules stained with the antibody is exactly the same as that of mitochondria stained with DAPI. (I-K) ef2-null cells were double-stained with the anti-Dd-EF2H antibody (I) and MitoTracker Orange (K), as described in Materials and Methods. It is clear that both the stains are completely merged (J). Bars, 10 µm.