Supplemental Material

This article contains the following supporting material:

• Figure 1 - Endoplasmic reticulum and actin cytoskeleton organizations are not altered in cells lacking Mfb1p. Perinuclear endoplasmic reticulum, and actin patches and cables were visualized by expressing Sec63p-GFP (A) and Phalloidin staining (B), respectively, in wild-type (JSY7452) and mfb1Δ (JSY7453) cells. Bar, 5 μm.

• Figure 2 - Mitochondrial membrane ultrastructure is not altered in cells lacking Mfb1p. Transmission electron microscopy was performed essentially as described previously (Rieder et al., 1996. Mol. Biol. Cell 7: 985-999) using wild-type (JSY7452) (A and B) and mfb1Δ (JSY7453) (C and D) cells grown in rich glycerol media. Bar, 2 μm.

• Figure 3 - Mitochondrial fission proteins localize properly in cells lacking Mfb1p. Wild-type (JSY7452) and mfb1Δ (JSY7453) cells expressing Dnm1p-GFP, GFP-Fis1p, or GFP-Mdv1p and a mito-RFP are shown. Bar, 5 μm.

• Figure 4 - Mitochondrial inheritance is not affected in the absence of Mfb1p. Mitochondria in wild-type (JSY7452) and mfb1Δ (JSY7453) cells were visualized using mito-GFP. Large- and small-budded cells containing mitochondria in the bud were scored (n = 300).

• Figure 5 - Skp1p localization patterns in budded cells. A skp1Δ mutant expressing Skp1p-3HA (Y94) and a wild-type strain expressing Skp1p (Y14) were subjected to DAPI staining and indirect immunofluorescence using anti-HA primary and FITC-conjugated secondary antibodies. The percentage of dividing cells with asymmetric Skp1p-HA localization in buds (I) or mother cells (II), uniform Skp1p-HA localization in mother cells and buds (III), or displaying other Skp1p-HA localization patterns (IV) is shown (n = 170). Bar, 5 μm.

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