Triglyceride Quantification in Tissue Homogenates

Tissue were homogenized and diluted in saline at a final concentration of 50 mg/mL as previously described^{61 (link), 65 (link)}. Homogenates were diluted (1:1) in 1% deoxycholate (Sigma-Aldrich) and incubated at 37 °C for 5 min. For triglyceride measurements, samples were diluted 1:100 in the reagent (Infinity™ Triglycerides Liquid Stable Reagent, Thermo Fisher Scientific) and incubated for 30 min at 37 °C. The resulting dye was measured based on its absorbance at 550 nm with a Sunrise ELISA plate reader (Tecan, Männedorf, Switzerland). Concentrations were determined compared with a standard curve of triglycerides (Infinity™ Triglycerides Standard, Thermo Fisher Scientific). The protein content of the preparations was measured by the Bradford method, using BSA (Sigma-Aldrich) as standard. All assays were performed in duplicate.