Journal: PLoS ONE

Article Title: Coxsackievirus A16 Elicits Incomplete Autophagy Involving the mTOR and ERK Pathways

doi: 10.1371/journal.pone.0122109

Figure Lengend Snippet: (A) Western blotting (WB) analysis of LC3 protein expression in HeLa cells infected with Coxsackievirus A16 (CA16). Cells were infected with CA16 at an MOI of 1. After 1 h of virus absorption at 37°C, the cells were further cultured in maintenance medium. Cells were harvested at the indicated time points and blotted with anti-LC3B and Vp1 antibodies; the results were compared to uninfected control cells. (B) GFP-LC3 dots and viral proteins expression were visualized via confocal microscopy. HeLa cells were transfected with GFP-LC3 plasmid for 24 h, followed by CA16 infection(MOI = 1) or treated with Rapamycin and the GFP-LC3 aggregations in the cells were assessed via confocal microscopy. The localization of virus was determined by indirect immunofluorescence with a human anti-CA16 sera and TRITC-conjugated anti-human IgG. Representative images are shown. Scale bar, 8μm. (C) Autophagic vacuoles were detected via transmission electron microscopy (TEM). HeLa cells infected with CA16 (MOI = 1) were processed and analyzed at 24 hpi for the accumulation of autophagosome via electron microscopy. White arrows indicate representative autophagosomes. (c) represents the higher-magnification views of (b).

Article Snippet: TRITC-conjugated goat anti-human antibody was obtained from Proteintech Group.

Techniques: Western Blot, Expressing, Infection, Virus, Cell Culture, Control, Confocal Microscopy, Transfection, Plasmid Preparation, Immunofluorescence, Transmission Assay, Electron Microscopy