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. Author manuscript; available in PMC: 2016 Sep 1.
Published in final edited form as: Biochim Biophys Acta. 2015 May 23;1851(9):1227–1239. doi: 10.1016/j.bbalip.2015.05.003

Fig. 5. Expression of Coq9-ts19 polypeptide affects steady state levels of other yeast Coq polypeptides in response to different growth temperatures.

Fig. 5

The mitochondria of W3031B (WT) and W303ΔCOQ9 harboring the temperature-sensitive plasmid TS19 (Δ9:TS19) were isolated after yeast were grown for 18.5 hours at either 25°C or 37°C. Mitochondria were also isolated from the null control strains W303ΔCOQ9 (Δ9), W303ΔCOQ7 (Δ7), and W303ΔCOQ4 (Δ4) after yeast were grown overnight at 30 °C. Purified mitochondria (15 μg protein) were separated by SDS-PAGE and analyzed by immunoblot. Immunoblots were performed with antibodies against the designated polypeptides: Coq4, Coq5, Coq6, Coq7, Coq9 and Atp2. Images presented within a given panel were derived from the same gel, and thus the band intensities corresponding to the same antisera can be directly compared. Densitometry quantification was conducted with the software ImageJ (NIH) and plotted as graphs. The signals of COQ4, COQ5, COQ6, COQ7, and COQ9 were each normalized to the signals of ATP2. Normalized values were then compared to WT at 25 °C to get the relative percentage. The immunoblot depicted is representative of two independent blots performed with two different preparations of purified mitochondria. (A, B, D) Bars designate the average signal + S.D. (n=2); (C, E) Each bar represents one measurement (n=1) because signals for Coq6 and Coq7 were detectable at 37 °C in only one set of the immunoblots.