FIG 8.

Overexpression of s-Mgm1 enhances Fzo1-independent mitochondrial fusion. (A) Percentage of cells with merged signals in mitochondrial fusion assay and 95% confidence intervals are shown. Counted cell numbers are indicated. Fisher's exact test was performed. Asterisks indicate P < 0.01. (B) Lysates of cells expressing chromosomally encoded Mgm1-Flag were subjected to Western blotting, and proteins were detected using the indicated antibodies. Pgk1 was used as a loading control. (C) Lysates of Mgm1-Flag-expressing cells harboring empty vector, the short isoform of Mgm1 (s-Mgm1)-Flag-, and full-length Mgm1 (FL-Mgm1)-Flag expression plasmids were subjected to Western blotting, and proteins were detected using the indicated antibodies. Pgk1 was used as loading control. The ratios of s-Mgm1 to l-Mgm1 were calculated. (D) The indicated yeast cells were treated with Zymolyase 100T to digest cell wall. Spheroplasts were homogenized, and the resultant homogenates were fractionated by differential centrifugation. Total, total homogenate; PMS, postmitochondrial supernatant; Mito, rough mitochondria. Pgk1 was used as a cytosolic loading control, and Om45-GFP was used as a mitochondrial loading control. (E) The mitochondrial fusion assay was performed as in Fig. 7A. FL, full length. (F) Percentages of cells with merged signals in the mitochondrial fusion assay and 95% confidence intervals are shown. Counted cell numbers are also indicated. Fisher's exact test was performed. Asterisks indicate P < 0.01. n.s., not significant.