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. Author manuscript; available in PMC: 2016 Apr 1.
Published in final edited form as: J Cell Physiol. 2015 Apr;230(4):831–841. doi: 10.1002/jcp.24811

Figure 6. Overexpression of full length wt BAG3 stabilizes the interaction of Bcl2 and Beclin-1.

Figure 6

A. Co-immunoprecipitation (co-IP) of Beclin-1 and Bcl2 in HEK293T cells. Lysates from cells transfected with Bcl2, Beclin-1 and BAG3 expression constructs (200 μg total protein) were used in co-IP experiments using antibodies to the FLAG tag on Beclin-1 or normal mouse serum (NMS) instead of FLAG antibody as a control for specificity of the complexes. Precipitated complexes were subjected to western blot analysis using antibodies to detected GPF (for Bcl2), Beclin-1 or myc (for BAG3). Note that FLAG-Beclin-1 migrates slightly above the endogenous Beclin-1 exhibiting a double protein band. B. Western Blot for Bcl2, Beclin-1 and BAG3 expression in HEK293T cells. Aliquots of HEK293T cell lysates (40 μg total protein) for each co-IP experiment were analyzed by western blot in parallel, resolved on the same SDS-PAGE, and probed against the same antibodies. GAPDH antibody was used on the same membrane as a loading control (Figure 6B, bottom panel).