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. 2024 Apr 9;13(8):655. doi: 10.3390/cells13080655

Figure 8.

Figure 8

VPS27 deletion in sit4∆ cells leads to a decrease in Atp2 and Cox2 levels, but it does not affect mitophagy or mitochondrial biogenesis. (A) Mitophagy is induced in sit4Δ cells in a Vps27-independent manner. Cells expressing pYX242-mtPHO8 were grown to Log and PDS (24 h after Log) phases and mitophagy was determined by measuring the alkaline phosphatase activity. Values are the mean ± SD (n = 4); * p ≤ 0,05, ** p ≤ 0.01, *** p < 0.001; one-way ANOVA. (B) Cells were grown to Log and PDS phases and the levels of mitochondrial proteins (Por1, Atp2, and Cox2) were assessed by immunoblotting. Hxk2 was used as a loading control. A representative blot is shown (n = 3). (C) Relative amount of Por1 (a mitochondrial outer membrane protein) to Hxk2 is shown as an indicator of mitochondrial mass. Values are the mean ± SD; ** p ≤ 0.01, *** p < 0.001; one-way ANOVA. (D,E) Quantification of the ratio Atp2/Por1 (D) and Cox2/Por1 (E). Values are the mean ± SD; * p ≤ 0.05, ** p ≤ 0.01; one-way ANOVA. (F) Analysis of mtDNA/nuclear DNA by qPCR. COX1 and COX3 were used as reference for mtDNA and ACT1 for nuclear DNA. Rho0 cells were used as control. Values are the mean ± SD (n = 4); **** p < 0.0001; one-way ANOVA.