FIG 4.

NCL is required for RABV infection. (A) U-373-MG cells transfected with Scr (control) or NCL-targeting siRNA (NCL1 [29]) were infected with CVS11 RABV at an MOI of 0.5 or 5, or mock infected, for 24 h. Cell lysates were then analyzed by IB using antibodies against NCL (rabbit antibody ab22758; Abcam), P-protein (mouse antibody 25C2 [39]), and tubulin (mouse antibody N3656; Amersham) and fluorescently conjugated anti-rabbit and anti-mouse IgG secondary antibodies. The blot shown is representative of 2 assays. (B) Cell lysates from the assay represented in panel A were analyzed by IB using antibody against RABV N protein (62B5) (40). (C) Viral titers were determined in supernatants from RABV-infected cells transfected with Scr or NCL1 siRNA (means ± standard deviations [n = 2] from 2 assays).