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. Author manuscript; available in PMC: 2013 Mar 2.
Published in final edited form as: Cell. 2012 Mar 2;148(5):988–1000. doi: 10.1016/j.cell.2012.01.038

Figure 1. Microsomes Restore Mitochondrial Sensitivity to Direct Activator Stimulation.

Figure 1

(A) HM and mitochondria were treated with C8-BID (0.5, 1, and 5 nM).

(B) The HM fraction was isolated, and the μS and mitochondrial (Mito) fractions were purified. All three fractions were subjected to western blot. In parallel, each fraction was incubated with 10 nM C8-BID for 1 hr at 37°C, washed, and subjected to western blot.

(C) Same as in (A), but treatments included BID BH3 and BIM BH3 (1, 5, and 20 μM).

(D) HM (50 μg) and mitochondria (50 μg) were treated as indicated (C8-BID, 1 nM; BID BH3, 5 μM; BCL-xL, 500 nM; μS, 5, 25, and 50 μg).

(E) HM, mitochondrial, and μS fractions were subjected to western blot.

(F) HM, mitochondrial, and μS fractions were treated with limited proteolysis, lysed, and subjected to western blot.

(G) Purification scheme to identify SMPD3 and SMPD4.

See also Figure S1.