Figure 6. LIR motifs are required for interaction of PML and LC3.

(a) Alignments of a portion of PML protein sequence across several mammalian species showed two conserved LIR motifs as indicated in the rectangles. Two potential LIR motifs were respectively mutated into the mutant form AXXA, designated mutant 1 (Mut1) and mutant 2 (Mut2). (b) HEK293T cells were transiently transfected with the indicated plasmids and Co-IP was performed with Flag antibody. The indicated proteins were analyzed by immunoblotting. 10% cell lysates (input) was used as a positive control. DM represents double mutant PML plasmid with the mutated sites of both mutant 1 and mutant 2. These experiments were repeated for three times and similar results were obtained.