FIG 5.

GBP-1 is necessary to mediate remodeling of the actin cytoskeleton induced by IFN-γ. (A) HeLa cells were transfected with either control siRNA or GBP-1 siRNA, as indicated. After 6 h, the cells were either left untreated or treated with IFN-γ (100 U/ml, 24 h). Next, 10 μg of protein lysate was separated via 10% SDS-PAGE and analyzed via Western blotting. GBP-1, actin, and GBP-2 (used as an siRNA specificity control) were detected using specific antibodies. (B) HeLa cells were transfected and treated with IFN-γ as described in the legend to panel A. The cells were stained for GBP-1 following antigen target retrieval (pH 9). The nuclei were counterstained with Draq5. (C) HeLa cells were treated as described in the legend to panel A and subsequently stained for actin. Asterisks, organized, filamentous actin cytoskeletons; arrowheads, disruption of the filamentous actin organization following IFN-γ treatment. The nuclei were counterstained with Draq5. All results were independently reproduced by a second researcher. Bars = 25 μm.