The accumulation of mutated NS1 protein within lysosomes does not involve autophagy. Vero cells were infected with either NS1F, NS1F-ELCmut, or mock infected with media, and cells were lysed in RIPA buffer 24 h and 48 h p.i. Equivalent amounts of total protein were separated by SDS-PAGE and analysed by Western blot. The active (LC3-II) and constitutive (LC3-I) forms of LC3 were detected using anti-LC3B polyclonal antibody. NS1 was detected using anti-FLAG monoclonal antibody. Representative of three independent experiments.