Fig 7. RABV P-protein antagonism of STAT3 is restored in U3A cells by STAT1 expression.

(A) 2fTGH and U3A cells co-transfected to express the indicated proteins were treated with or without OSM (10 ng/ml, 30 min) before fixation, immunofluorescent staining for STAT3 (blue, Alexa Fluor 647) and analysis by CLSM, as described in the legend to Fig 1. Representative images are shown. Arrowheads indicate cells with detectable expression of transfected proteins. Scale bars, 30 μm. (B) Images such as those shown in A were analysed to calculate the Fn/c for STAT3 (mean ± SEM; n ≥ 40 cells for each condition; results are from a single assay representative of three independent assays). Statistical analysis used Student’s t test. ****, p < 0.0001; NS, not significant. (C) U3A cells stably transfected with plasmid for tetracycline-inducible expression of STAT1 were cultured with or without 1 μg/ml tetracycline (Tet) to induce STAT1 expression and transfected to express GFP or GFP-CVS-Pwt. Cells were treated 24 h post-transfection with or without OSM before immunoprecipitation of GFP and analysis by IB, as described in the legend to Fig 3. Results are representative of two independent assays.