Figure 3.

Mitochondria functioning is changed upon GDAP1 expression in yeast. (A) Overnight cultures of wild-type yeast cells harboring plasmid bearing mitochondrially targeted green fluorescent protein (mt-GFP) and empty vector ([-]) or plasmid with full-length GDAP1 were shifted to glycerol-containing medium for 4 h to induce expansion of mitochondria. Mitochondrial morphology was observed using fluorescent microscopy. The described phenotypes were counted. Error bars represent standard deviation for three independent biological experiments. (B) PTY44 strain transformed with empty vector ([-]) or GDAP1 gene variants (as indicated) was incubated for 2 days in glycerol-containing medium and then the cells were counted, and dilutions of every culture were plated on YPD or SC-trp medium. After 3 d of incubation at 28 °C the number of colony-forming units (CFUs) was calculated. The fraction of SC-trp-growing cells in every culture were normalized to PTY44 with empty vector ([-]). Error bars represent standard deviation for four experiments. (C) Wild-type yeast strain harboring empty vectors ([-]) or plasmids with GDAP1 alleles were grown overnight in SC gly-leu medium. The cultures were diluted, and cells were plated on YPG + 0.2% glucose and incubated for 3 d at 28 °C. CFUs were counted and the percentage of small colonies was calculated. Error bars represent standard deviation for four experiments.