Figure 2

Redistribution of GFP-LC3 autophagy marker in ChikV-infected HEK.293 cells. (A) GFP-LC3-HEK.293 stable cells cultured on glass coverslips were incubated from 2 h to 48 h with ChikV (MOI = 1) or mock infected (Ct) and analysed by fluorescence microscopy to determine GFP-LC3 distribution. The status of cells regarding infection was determined by detection of ChikV antigens using a monoclonal antibody directed against the E1 glycoprotein and Alexa Fluor 594 (red)-conjugated secondary reagent. Nuclei were stained with DAPI. Magnification = 200× (upper panel), magnification = 600× (lower panel) (B) GFP-LC3 positive cells were defined as cells that display more than 5 dots in the cytoplasm. Numbers of GFP-LC3 positive cells were counted on more than 100 cells. (C) For each positive cell, the number of GFP-LC3 dots was counted. These results are expressed as mean values obtained from three independent experiments +/- standard errors.