Research Paper Volume 8, Issue 12 pp 3255—3271

Figure 5. The function of Slm35 is necessary for the regulation of general and mitochondrial selective autophagy. (A) Relative survival under heat shock conditions of wild-type (WT), single mutants Δslm35, Δatg1, Δatg4, Δatg6, Δatg17, and Δatg21, or double mutants Δatg1Δslm35, Δatg4Δslm35, Δatg6Δslm35, Δatg17Δslm35, and Δatg21Δslm35. Cells were grown on SCD liquid medium during three days before applying a thermal shock of 20 min at 55 ^oC. Aliquots were taken immediately after the stress to construct YPD growth curves to calculate the relative survival. (B) Overnight YPD cultures of wild-type or Δslm35 cells harboring the fusion protein Idh1-GFP were shifted to YPL medium to an OD₆₀₀=0.1 to induce mitophagy. Every 24 hours an aliquot of the culture was taken and whole cell extracts were analyzed by Western blot. Mitochondrial Idh1-GFP runs as a discrete band at around 60 kDa, vacuolar free GFP runs at 25 kDa. G6PDH was used as loading control. Signals from three independent experiments were quantified by densitometry.