Fig. 6
From: A nile red fluorescence assay for LC3 autophagy protein binding to lipid bilayers
Effect of SM hydrolysis by SMase on LC3 protein binding to lipid bilayers. Data for LC3A, LC3B, and LC3 proteins are given. Top row, maximum observed values of % ROIR decrease. Middle row, slopes of the % ROIR vs. LC3 concentration curves. Bottom row, % bound protein, measured with an ultracentrifugation-flotation method10. Bilayer lipid compositions were: 1, eSM: DOPE (50:50); 2, eSM: DOPE: Cer (40:50:10); 3, eSM: DOPE (50:50) + SMase; 4, eSM: DOPE: CL (33:33:33); 5; eSM: DOPE: CL: Cer (23:33:33:10); 6, eSM: DOPE: CL (33:33:33) + SMase. SMase was allowed to act for 1 h, a time period in which released Cer was about 10% of the total lipid. Average values ± S.D., n = 3. Statistical significance was estimated according to the Student’s t-test, p values are indicated for each pair of significantly different data: ** p < 0.01, *** p < 0.001.
