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. 2010 Sep 24;18(3):439–451. doi: 10.1038/cdd.2010.114

Figure 4.

Figure 4

INrf2 decreases Bcl-2:Bax interaction. (a) IP/immunoblot analysis of Bcl-2 interaction with INrf2 and Bax. INrf2-293 cells were treated with tetracycline (0.5 μg/ml) for different time periods to express Flag-INrf2. The cells were harvested and 1 mg of cell lysates was immunoprecipitated with anti-Flag or anti-Bcl-2 antibody and immunoblotted with anti-Bax, anti-Bcl2, and anti-Flag antibody. (b) Immunoblot analysis of the effect of overexpression of Flag-INrf2 on Bcl-2 and Bax. Hepa-1 cells were co-transfected with increasing concentrations of Flag-INrf2, along with the same concentration of Bcl-2-V5, and cell lysates were immunoblotted with indicated antibodies (input panel). (c–e) IP/immunoblot analysis of Bcl-2 interaction with INrf2 and Bax. A volume of 1 mg of cell lysates from transfected Hepa-1 cells in ‘b' was immunoprecipitated with either anti Flag-INrf2 (c) or anti-V5 (d) or anti-Bax (e) antibodies and immunoblotted with indicated antibodies. (f) The relative interaction of Bcl-2-V5:Bax and Bcl-2-V5:Flag-INrf2 was quantified from three experiments and plotted as mean±S.D.