Figure 2. The levels of mitochondrial... | Rockefeller University Press

Figure 2.

$The levels of mitochondrial proteins are post-transcriptionally affected upon eIF5A depletion. (A and B) Polysome profiles were obtained for wild-type and tif51A-1 strains cultured in SGal at 25°C (A) or 37°C (B) for 4 h. A representative profile is shown from three independent experiments (left). The average polysome and monosome area is represented for each strain at both temperatures (right). (C) The RNA from individual fractions of the polysome profiles was extracted and the mRNA levels were analyzed by RT-qPCR using gene-specific primers in the corresponding sections at restrictive temperature. Fold change (FC) between the heavy and the low polysomal sections are presented. (D) Wild-type strain and tif51A-1 expressing nLuc, Cyc1-nLuc, Cox5a-nLuc, or Sdh2-nLuc (from left to right) reporters were cultured in YPD until early exponential phase and then transferred to 25°C or 37°C for 4 h. After addition of doxycycline to induce luciferase expression, the luminescence levels generated by the nLuc 60 min after the addition of the furimazine substrate were measured. Data information: In A–D, results are presented as average ± SD from three independent experiments. The statistical significance was measured by using a two-tailed paired Student’s t test. *P < 0.05, **P < 0.001, ***P < 0.001. n.s means no significant differences.$

The levels of mitochondrial proteins are post-transcriptionally affected upon eIF5A depletion. (A and B) Polysome profiles were obtained for wild-type and tif51A-1 strains cultured in SGal at 25°C (A) or 37°C (B) for 4 h. A representative profile is shown from three independent experiments (left). The average polysome and monosome area is represented for each strain at both temperatures (right). (C) The RNA from individual fractions of the polysome profiles was extracted and the mRNA levels were analyzed by RT-qPCR using gene-specific primers in the corresponding sections at restrictive temperature. Fold change (FC) between the heavy and the low polysomal sections are presented. (D) Wild-type strain and tif51A-1 expressing nLuc, Cyc1-nLuc, Cox5a-nLuc, or Sdh2-nLuc (from left to right) reporters were cultured in YPD until early exponential phase and then transferred to 25°C or 37°C for 4 h. After addition of doxycycline to induce luciferase expression, the luminescence levels generated by the nLuc 60 min after the addition of the furimazine substrate were measured. Data information: In A–D, results are presented as average ± SD from three independent experiments. The statistical significance was measured by using a two-tailed paired Student’s t test. *P < 0.05, **P < 0.001, ***P < 0.001. n.s means no significant differences.