Figure 3.

Expression of Dnm1p causes fragmentation of mitochondria in dnm1Δ fzo1Δ cells. dnm1Δ fzo1Δ cells carrying pCOX4-GFP (pHS12) and pGAL1-DNM1-HA (pHS15) were pregrown in raffinose medium, centrifuged and resuspended to an OD₆₀₀ of 0.2 in galactose medium (SGS) to induce Dnm1p-HA expression. Cells were examined for mitochondrial shape (n = 100) or used to prepare total protein at the indicated timepoints. (A) Mitochondrial shape was classified into the following four groups: tubules (○), partially fragmented tubules (□), fragments (▴), and other (⋄). (B) Total protein was extracted as described ( Yaffe and Schatz 1984) and subjected to Western blot analysis using antibodies to the HA epitope ( Field et al. 1988), or hexokinase (Davis, A., unpublished data) followed by chemiluminescence (Pierce). Relative amounts of Dnm1p-HA (+) were quantitated and plotted in A.