Fig. 2

Loss of PFD2 and PFD5 results in defective mitochondria network. A, D Yeast cells that were transformed with a plasmid that harbored mitochondrial-targeted GFP (mtGFP) were grown at 25 °C on selective synthetic medium that contained glycerol or glucose as indicated. Representative images are shown for the expression of mitochondrial GFP and images processed with the MINA plugin tool in ImageJ (see “Methods” section) of the same cell are shown below. Pink lines represent mitochondrial footprint generated by MINA plugin tool. Scale bar = 5 µm. B Schematic representation of features taken into account to quantify the number of mitochondrial networks shown in C and E. C, E Analysis of mitochondrial morphology, expressed as the number of network points. The data are expressed as the mean ± SEM. n = 3. ***p < 0.001, **p < 0.01, *p < 0.05. D–I Wildtype cells or cells that lacked PFD2 or PFD5 were transformed with an empty vector or a plasmid that harbored PFD2-Flag or Flag-PFD5, respectively. F Ten-fold dilutions of yeast cells of the indicated strains were spotted on selective medium plates that contained glycerol and 0.5% galactose. Cells were grown at 37 °C for 6 days. Experiments were performed in at least two biological repetitions. G, H Transformed yeast cells were grown in selective G, H glycerol- or I glucose-containing liquid medium to the logarithmic growth phase at 25 °C. Galactose (0.5%) was added to the cultures to induce expression from the plasmid, and cells were shifted to 37 °C for 4 h. Cell viability was assessed by staining with propidium iodide and analyzed by flow cytometry. The data are expressed as the mean ± SEM. n = 3. ***p < 0.001; ns, not significant. WT, wild type; EV, empty vector