Illuminating the Sites of Enterovirus Replication in Living Cells by Using a Split-GFP-Tagged Viral Protein

van der Schaar, H. M.; Melia, C. E.; van Bruggen, J. A. C.; Strating, J. R. P. M.; van Geenen, M. E. D.; Koster, A. J.; Bárcena, M.; van Kuppeveld, F. J. M.

Western blot analysis. Proteins were separated with 10% gradient PAGE using the Gradi-Gel gradient analysis kit (Elpis Biotech). Samples were transferred to nitrocellulose membranes (Bio-Rad). After the membrane was cut at the 15-kDa band of the marker, one membrane (proteins larger than 15 kDa) was incubated with primary antibodies against mouse monoclonal anti-␤-actin (Sigma) and rabbit polyclonal anti-GFP described previously (44) , which also recognizes GFP(S1-10). The membrane with proteins smaller than 15 kDa was incubated with rabbit polyclonal anti-3A described previously (44) . Secondary antibodies included IRDye 680-conjugated goat anti-mouse or IRDye 800-conjugated goat anti-rabbit (LI-COR). Images of blots were acquired with Odyssey imaging system (LI-COR).