## 4 Materials and Methods
### Preparation and Analysis of Lipids  
The lipids were extracted from the mitochondria by the Nichols method described in [53], which involved extraction with isopropanol and the isopropanol-chloroform mixture (1:1 and 1:2) at 70 \({}^{\circ}\)C, evaporation in a rotary evaporator, and extraction of the residue with chloroform-methanol (1:1) supplemented with 5% sodium chloride solution and water to remove water-soluble substances. After separating the mixture with a vortex, we dried the chloroform layer by passing it through water-free sodium sulphate, evaporated, and desiccated with a vacuum pump. The resulting pellet dissolved in a small amount of chloroform-methanol (2:1) was stored at -21 \({}^{\circ}\)C. The composition of storage lipids was assayed using an ascending thin layer chromatography on glass plates with silica gel 60 (Merck KGaA, Darmstadt, Germany). To separate storage lipids, the hexane: sulphuric ether: acetic acid (85:15:1) system [54] was used. To separate PLs and SLs SI60 Silica thin layer chromatography plates were activated and developed in two dimensions, first with chloroform/methanol/water (65/25/4, by volume) and second with chloroform/acetone/methanol/acetic acid/water 50/20/10/10/5, by volume) [55]. The lipids (100-200 ug) were applied to a plate. Lipid quantities were determined using the following standards: PC (Sigma, Saint-Luis, MO, USA) for PLs, a glycoceramide mixture (Laordan, Solna, Sweden) for SLs, and ergosterol (Sigma, Saint-Luis, MO, USA) for St. Samples of glycoceramides (5 and 10 ug) and PC (10 and 20 ug) were applied on the plates in the second direction. To develop the stains, the chromatograms were sprayed with 5% sulfuric acid in ethanol, followed by heating up to 180 \({}^{\circ}\)C. Quantitative densitometric analysis of the lipids was performed using the Dens software package (Lenkhrom, Sankt Peterburg, Russia) in the linear approximation regime using the calibration curves constructed with the standard solutions. Lipid data are presented in ug/g mitochondrial protein. PLs were identified using individual markers and qualitative tests for amino groups (with ninhydrin), choline-containing phospholipids (with the Dragendorff reagent), and glycolipids (with \(\alpha\)-naphthol). Neutral lipids were identified with individual markers for MAGs, DAGs, and TAGs, St (ergosterol), FFAs, and hydrocarbons (Sigma, Saint-Luis, MO, USA). SLs were detected in the glycolipid fraction by the saponification method [56].  
To assess the FAs composition of PLs, separate PLs were isolated using TLC with two plates, eluted with chloroform/methanol (1/1, _v/v_) for a night. Then, the supernatant was decanted, evaporated, 1 mL toluene and 2 mL of 2.5% H\({}_{2}\)SO\({}_{4}\) dissolved in methanol and kept for two hours at +70 \({}^{\circ}\)C. FAs methyl ethers were extracted with hexane, dried, and analyzed by a Kristall 5000.1 gas chromatograph (Chromatek, Moscow, Russia) using an Optima-240 (60 m \(\times\) 0.25 mm) capillary column (Macherey-Nagel GmbH and Co., Dueren, Germany). The temperature program was set from +130 to +240 \({}^{\circ}\)C. Eluting FAs were identified using the Supelco 37 Component FAME Mix (a mixture of FA methyl esters) (Supelco, Bellefonte, PA, USA). The degree of unsaturation was calculated by the equation: \(\Delta\)/mole = 1.0 \(\times\) (% monoene)/100 + 2.0 \(\times\) (% dienes)/100 + 3.0 \(\times\) (% trienes)/100 [57].