The late wave of autophagy induced by attenuated MeV is productive.

<p>(A) HeLa cells were infected with Sch-MeV (MOI 1). At the indicated time points post infection, cellular p62 expression was determined by western blot. (B) HeLa cells were infected with Ed-MeV (MOI 1). At the indicated time points post infection, cellular p62 expression was determined by western blot. The graph represents the intensity of p62 over actin expression normalized to the control condition (non infected). Error bars, mean ± MD of two independent experiments. (C) HeLa cells were infected with Ed-MeV (MOI 3). 24 h p.i., cellular p62 expression was determined by western blot. Representative results are shown and are accompanied by a graph representing the intensity of p62 over actin expression normalized to the control condition (non infected). (D) mRFP-GFP-LC3 HeLa cells were infected with Ed-MeV (MOI 3) or treated with 250 nM rapamycin (Rapa) or 75 µm Chloroquine (CQ) during 2 h. 24 h p.i., the total number of autophagic vesicles (mRPP+) and the number of autophagosomes (mRFP+/GFP+) were assessed by confocal microscopy. The number of autolysosomes was determined by subtracting the number of autophagosomes (mRFP+/GFP+) from the total number of vesicles (mRPP+). Representative profiles are shown and are accompanied by a graph representing the number of autophagic vesicles (white), autophagosomes (yellow) and autolysosomes (red) per nuclear profile for each condition. (E–F) HeLa cells were treated with the indicated si-RNA for 48 h and infected with Ed-MeV (MOI 3 (E) or MOI 0.1 (F)). 24 h (E) or 48 h (F) post-infection, cells were lysed and anti-N and anti-P western blot were performed to reveal MeV-N and MeV-P, respectively. Representative results are shown and are accompanied by a graph representing the intensity of MeV-N or MeV-P expression over cellular actin normalized to the control condition. (C–F) : error bars, mean ± SD of three independent experiments. Student's t test; ***p<0.005; **p<0.01; *p<0.05; #p>0.05.