Figure 3.

NF-κB-independent role of T6BP and NDP52 in MeV replication. (A) p65/RelA-expressing HeLa cells and shControl-expressing HeLa cells were transfected with the indicated siRNAs for 48 h, then lysed, and the expression of relevant proteins was probed by Western blot; (B) p65/RelA-expressing HeLa cells and shControl-expressing cells were infected with MeV (MOI 0.1). 48 h post infection, infectious virus particles were titrated by a plaque assay; (C) Cells from (B) were lysed 48 h post infection. Expression of measles virus N and P proteins were assessed by Western blotting. Representative results from shp65#1 are shown and are accompanied by a graph representing the intensity of MeV-N and MeV-P expression over Actin normalized to shControl-expressing cells condition. Means ± SD of four independent experiments are represented (two with the shp65#1 cell line and two with the shp65#2 cell line); (D) p65/RelA-expressing HeLa cells and shControl-expressing HeLa cells were stained for CD46 expression and analyzed by flow cytometry; grey histograms = isotype control, white histograms = CD46 labelling. (E–G) p65/RelA-expressing HeLa cells were treated with indicated siRNAs for 48 h; (E) Cells were lysed and the expression of relevant proteins was probed by Western blotting. Results regarding cell line shp65 #1 are represented. Similar results were obtained with shp65 #2. Cells were infected with MeV (MOI 0.1) and 48 h post infection, infectious virus particles were titrated by a plaque assay (F) or lysed; (G) Expression of measles virus MeV-N and MeV-P proteins was assessed by Western blotting. Representative results are shown and are accompanied by a graph representing the intensity of measles proteins expression over Actin normalized to control siRNA condition; (B,F) Means ± SD of one representative experiment out of two independent ones carried out with each shp65/RelA-expressing cell line in duplicates; (G) Means ± SD of four independent experiments.