The ultrastructural localization of LC3B was examined using zebrafish larvae, employing the post-embedding method as described previously49,50. Small larvae specimens embedded in LR White Resin, prepared as semi-thin sections, were used. The RFP antibody was used for the detection of mCherry protein, because it reacts with RFP and other RFP variants, such as mCherry. Ultra-thin sections (70 nm thick) were cut, incubated with a rabbit polyclonal LC3B antibody (1:300) and a mouse monoclonal RFP antibody (1:100) for 2 h at 24 °C , and reacted with 10-nm gold colloidal particle-conjugated anti-rabbit IgG (EMGFAR10; British BioCell International, Cardiff, UK; 1:30) and 5-nm gold colloidal particle-conjugated anti-mouse IgG (EMGMHL5; British BioCell International; 1:30). Finally, the sections were stained with lead citrate and examined using a JEM-1400 electron microscope at 80 kV (JEOL, Tokyo, Japan).

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