Mouse anti his

Skin biopsies were fixed in 4% paraformaldehyde (Sigma-Aldrich, St. Louis, MO, USA at 4 °C overnight. The next day, skin biopsies were buffered in a 30% sucrose solution (Sigma-Aldrich, St. Louis, MO, USA) and stored at 4 °C. For sectioning, biopsies were embedded in O.C.T. compound (Sakura, Tokyo, Japan) and sectioned at a thickness of 15 μm using an OTF cryostat (Bright Instruments, Cambridge, UK). Sections were stained with unconjugated primary goat anti-FLAG (QED Bioscience, San Diego, CA, USA), mouse anti-His (Abcam, Cambridge, UK), or rabbit anti-Myc (Abcam, Cambridge, UK) antibodies. Sections were then stained with donkey anti-goat AF488 (Abcam, Cambridge, UK), donkey anti-rabbit AF555 (Life technologies, Carlsbad, CA, USA), and goat anti-mouse AF647 (Life technologies, Carlsbad, CA, USA) conjugated secondary antibodies, respectively. An additional stain, Hoechst 33342 (Life Technologies, Carlsbad, CA), was used to visualize nuclei. The slides were then mounted with Fluoromount (eBioscience, San Diego, CA, USA) and viewed by fluorescence microscopy using an Olympus BX51 with a U-TV1X-2/U-CMAD 3 combo camera for photo acquisition (Olympus, Melville, NY, USA). MagnaFire software was used to acquire the images.

The following primary antibodies were used: rat anti-atg8a (a gift from G. Jusász), rabbit anti-M1BP^{22 (link)}, mouse anti-c-Myc (9E10; Santa Cruz Biotech; SC-40), mouse anti-His (Abcam; ab9136) and rabbit anti-LC3A (Cell Signaling Techology).