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. 2015 Nov 30;291(4):1957–1973. doi: 10.1074/jbc.M115.668228

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FIGURE 2. — SIRT3 is associated with mitochondrial CerS1, -2, and -6. SIRT3 association with CerS1, CerS2, and CerS6 was detected in reciprocal immunoprecipitation (IP) experiments. Mitochondria were immunoprecipitated with anti-CerS6 antibody (Novus) and were probed using anti-SIRT3 (Cell Signaling), anti-CerS1 (Santa Cruz Biotechnology), anti-CerS2 (Santa Cruz Biotechnology), anti-CerS4 (Santa Cruz Biotechnology), or anti-SIRT5 (Cell Signaling) antibodies (A); with anti-SIRT3 antibody (Cell Signaling) and probed using anti-CerS1 (Santa Cruz Biotechnology), anti-CerS2 (Novus), anti-CerS4 (Santa Cruz Biotechnology), or anti-CerS6 (Novus) antibodies (B); with anti-SIRT5 antibody (Santa Cruz Biotechnology) and probed using anti-CerS1 (Santa Cruz Biotechnology), anti-CerS2 (Santa Cruz Biotechnology), or anti-CerS6 (Abcam) antibodies (C); and with anti-CerS2 antibody (Novus) and probed using anti-SIRT3 (Santa Cruz Biotechnology), anti-CerS1 (Santa Cruz Biotechnology), anti-CerS4 (Santa Cruz Biotechnology), or anti-CerS6 (Abcam) antibodies (D). Input load: 20 μg/lane. As a control, the same immunoprecipitation procedure was performed except for primary antibody application (IgG).