Figure 5

Interaction of PABPC1 with M2-1 expressed either in an infectious context or alone. (a) HEp-2 cells were infected with either RSV-M2-1-GFP or RSV-GFP (control) for 14 h and then subjected to co-IP with GFP antibody. RNAse (RNAse A or broad-spectrum nuclease) was added or not during the lysis step as indicated. After SDS-PAGE, immunoblot probing was then performed to detect the M2-1-GFP, GFP, PABPC1 and wild-type M2-1 proteins in the whole cell lysate (input) and in the bound fraction (bound). Full blots are available in Supplementary Fig. S5. Representative images from six to seven independent experiments are shown. (b) HEp-2 cells were transfected with either M2-1-GFP or GFP (control) for 24 h, and then subjected to co-IP with an anti-GFP antibody. RNAse A was added or not during the lysis step as indicated. Immunoblot probing was then performed to detect the M2-1-GFP, GFP and PABPC1 proteins. Full blots are available in Supplementary Fig. S7(a–c). Representative images from three independent experiments are shown. (c) HEp-2 cells were infected with wild type RSV for 14 h, and then subjected to co-IP with an antibody directed against either PABPC1 or a non-relevant protein (IMPDH2). RNAse A was added during the lysis step. M2-1 and PABPC1 were then detected by Western blotting. Full blots are available in Supplementary Fig. S7(d–e). Representative images from two independent experiments are shown.