Figure 1. The two types of mRFP-GFP-LC3 puncta have different dynamics in living cells. (A) Schematic diagrams of LC3 associated with the autophagosomal structures and protein aggregates. Left panel: LC3 is bound to both sides of the autophagosomes. Immediately before and after fusion with the lysosomes, the outer membrane-bound LC3 is released from the autophagosomes, and the inner membrane-bound LC3 is degraded in the autolysosomes. Right panel: During the formation of LC3-involved aggregate, the ubiquitinated proteins first interact with SQSTM1, then become protein aggregates through the oligomerization of SQSTM1. LC3/LC3G120A are recruited to the aggregates through their interaction with the LC3 interacting region (LIR) of SQSTM1. (B) FRAP analysis of the mRFP-GFP-LC3 puncta (arrows) in HBSS-starved HeLa cells. Scale bar: 5 μm.