Autophagic degradation of large T antigen is induced by Bag3.

<p>A. BSB8 cells were transfected with an expression plasmid encoding Bag3, and treated either with pepstatin A, 3-MA, or rapamycin. Whole cell protein extracts were prepared at 48 hour post-transfection, and Western blot was performed for the detection of T-Ag, and Bag3 proteins. Western blot analysis of the same extracts with anti-Grb2 antibody was used as loading control. B. Relative intensity of the band corresponding to T-Ag from Panel A is shown as a bar graph. C. BSB8 cells were either transfected with control empty plasmid (lane 1) or expression plasmid encoding Bag3 protein (lane 2). Protein extracts were prepared at 48 hour post-transfection, and Western blot analysis was performed for the detection of Large T, Bag3, Bag1, Hsp70, LC3, and Grb2 proteins. D. Relative intensity of the bands corresponding to LC3-II seen in Panel C were determined by densitometry and after normalization to that from Grb2, illustrated as a bar graph.