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For serum samples, 75 L of chloroform:methanol extraction solvent (v:v = 1:2) was added to 20 L of mouse serum, with the exception of the serum from WT, Gclc L-KO, Nrf2 L-KO, and L-DKO mouse serum, for which 20 L of mouse serum was combined with 180 L of chloroform:methanol extraction solvent (v:v = 1:2) containing internal standards at the final concentrations: 5 nM D7-Sphinganine (Avanti Polar Lipids Inc., Cat# 860658), 12.5 nM D3-Deoxysphinganine (Avanti Polar Lipids Inc., Cat# 860474), and SPLASH LIPIDOMIX (1:1000, Avanti Polar Lipids Inc., Cat# 330707)

After harvesting the cells by centrifugation at 4,000 g for 30 min, the cells were resuspended in lysis buffer (50 mM sodium phosphate, pH 8.0, 300 mM NaCl, 10 mM imidazole) and lysed using a sonication for 1 h with a 10-s pulse mode (10 s on and 10 s off) on ice