Fig. 4. Effect of overexpression of Gcs1p on mitochondrial morphology. Data are presented as in Fig. 3. (A) lysates of spheroplasts from gcs1 cells overexpressing wild-type recombinant Gcs1p under the control of the ADH1 promoter and grown in minimal medium were fractionated for analysis by immunoblotting. (B) and (C) indirect immunofluorescence staining of gcs1 cells overexpressing wild-type Gcs1p. (B) spheroplasts were reacted with purified anti-Gcs1p antibodies (b) or anti-porin antibody (c) or stained with H33258 (a). Arrows indicate perinuclear localization and arrowheads mitochondria. Panel C, spheroplasts were reacted with purified anti-Gcs1p antibodies (b) or anti-actin antibody (c) or stained with H33258 (a). Arrowheads indicate colocalization of Gcs1p and actin structures.