Figure 2. Primary neurons utilize STAT1-independent pathways during IFNγ-mediated viral control.

(A) NSE-CD46⁺ and NSE-CD46⁺/STAT1-KO hippocampal neurons were treated with IFNγ (100 U/ml) and infected with MV (MOI=1) for 24 or 48 h. Whole cell lysates were harvested, subjected to Western blot analysis, and quantified on ImageJ software, with MV signal normalized to GAPDH. The average MV signal is plotted with MV infection at 24h set to 100%, and error bars represent SEM (n=3). (B) Immunofluorescence analysis of MV antigen in CD46⁺ and CD46⁺/STAT1-KO neurons. Neurons were treated as in (A) and stained for MV antigen (red), MAP2 (green) and Hoescht (blue). Original magnification = 400X. (C) Primary CD46⁺ neurons were either treated with IFNγ 24h before infection (lane 2), coincident with infection (lane 3), or 24h post-infection (lane 4). Lysates were collected from infected (and corresponding uninfected) neurons, and blots probed for MV antigen and GAPDH as described. (D) CD46⁺ neurons were either treated with IFNγ (100 U/ml) or left untreated and 24 h later, infected with MV. 48 hpi, coverslips were collected and immunostained for MV antigens, and nuclei within immunopositive clusters counted. Clusters were “binned” into groups of 1–3, 4–6, 7–10 or greater than 10 (n=3). Standard deviations are shown from three such experiments.