Anti-ATG2A Blotting in Human Macrophages

For the anti-ATG2A blotting shown in Fig. 5, primary human macrophages were treated with IL-6 80 ng μl⁻¹ for 24 h before being lysed. The primary antibody used was anti-ATG2A pAb (MBL Life Science) at 1:400. Other western blots were performed with anti-Drosophila Atg8 (courtesy Katja Köhler (University of Zürich)57 (link) and G. Juhasz (Eötvös Loránd University)58 (link), both used at 1:200) and anti-α-tubulin (Developmental Studies Hybridoma Bank 12G10, used 1:10,000). Immunofluorescence on fly cells was performed with fluorescein isothiocyanate-labelled anti-M tuberculosis (Invitrogen PA1-28997, used 1:50) and rabbit anti-GFP (Invitrogen A-11122, used 1:100). Immunofluorescence on human cells was performed with anti-V5 from Novus Biologicals (NB 600-381), 1:400.