Journal: International Journal of Medical Sciences

Article Title: Parvovirus B19 Nonstructural Protein-Induced Damage of Cellular DNA and Resultant Apoptosis

doi:

Figure Lengend Snippet: PARP is active and necessary for efficient apoptosis in NS1-transfected cells. A. Immunoprecipitated GFP/NS1 protein was poly(ADP ribose)ylated, as shown by a band at 100 kd on the blot probed with anti-poly ADP ribose (PAR, left blot) antibodies. The blots were stripped and reprobed with anti-GFP (right), showing that PAR colocalizes with the GFP/NS1 fusion protein. GFP alone is not ribosylated, as evidenced by the lack of a PAR band corresponding to GFP at 37 kD. Blots shown are representative of three independent experiments. B. PARP activity is necessary for optimal NS1-induced apoptosis. Addition of the PARP inhibitor 5-aminoisoquinolinone (5AIQ) to GFP/NS1 expressing HepG2 cells reduced apoptosis by 57% ( p <0.003). Addition of 5-aminoisoquinolinone had no effect on the GFP transfected cells. N=3, error bars indicate the range of values.

Article Snippet: 25 μl of anti-GFP polyclonal antibody (Rockland, Gilbertsville, PA) were added and the mixture was allowed to bind for 14 hours at 4 o C in an end-over end rotator.

Techniques: Transfection, Immunoprecipitation, Activity Assay, Expressing