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. 2012 Jan 25;287(12):9112–9127. doi: 10.1074/jbc.M111.248906

FIGURE 4.

FIGURE 4.

Endogenous Bax does not co-IP with 14-3-3, Mcl-1, or Bcl-xL. 100 μg of cytosolic proteins of healthy and IL-3-deprived FDM (A) or FDC-P1 (B and C) cells were immunoprecipitated (IP) with 5 μl of the indicated antibody (Bax, N-Bax, 14-3-3, Mcl-1, Bcl-x). The IPs and resulting supernatants after IP (SN) were loaded on a 15% SDS gel and immunoblotted with anti-Bax (NT) or the indicated antibody. A, anti-N-Bax, anti-C-Bax, or anti-14-3-3 IPs are followed by anti-14-3-3 (upper row) or anti-Bax (lower row) Western blots. B, anti-N-Bax or anti-Mcl-1 IPs are followed by anti-Mcl-1 (upper row) or anti-Bax (lower row) Western blots. C, anti-Bcl-x or anti-N-Bax or anti-C-Bax IPs are followed by Bcl-x (upper row) or anti-Bax (lower row) Western blots. h, hours; Input, cytosol before IP; α-C, antibody against the C terminus of Bax (amino acids 184–192, supplemental Fig. S3); α-N, antibody against the N terminus of Bax (amino acids 2–14, supplemental Fig. S3). In B, the Ig heavy chains run just above Mcl-1. The molecular masses of the respective proteins are indicated.