Fig 9.
Epifluorescence analysis of proximity ligation assays. Cells were transduced with the NS5A-v1 lentivirus (A and B) or the mutated NS5A-v1 lentiviruses Lenti-NS5A-v1/NLSmut (C and D) and pLenti-NS5A-v1/D154E (E and F). In panels A, C, and E, proximity ligation was performed using a mouse anti-NS5A antibody (OBT 1222) and a rabbit anti-NS5A antibody (29) as positive controls to confirm the expression of NS5A in this transduced cells. In panels B, D, and F, proximity ligation was performed using a mouse anti-NS5A antibody (OBT 1222) and a rabbit anti-ASH2L antibody. This analysis confirmed the presence of a small proportion of NS5A in cell nuclei and indicated that NS5A translocation to the nucleus is reduced or abolished by impairing NS5A NLS or caspase cleavage.