Figure 4. Immunoblot analysis of hydrodynamically delivered, tagged Atg4b substrates.

Mice were injected with mammalian expression vectors containing tagged cDNAs for Lc3 (A), Gabarap (B), Atg8l (C), and Gate-16 (D) through the tail vein to achieve transient transgene expression in vivo. Liver protein extracts from injected WT and Atg4b^–/– mice were subjected to immunoblotting. Note that the absence of Atg4b impaired the processing of all of proteins, although its impact on Gate-16 was lower than in Lc3, Gabarap, and Atg8l. Unknown bands, which likewise corresponded to partial degradation products, are denoted by asterisks. β-Actin was used as a loading control.