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. Author manuscript; available in PMC: 2015 Mar 4.
Published in final edited form as: Cell Signal. 2010 Apr 18;22(9):1300–1307. doi: 10.1016/j.cellsig.2010.04.006

Fig. 6. CerS2, 5, and 6 form heterocomplexes.

Fig. 6

(A) Lysate was prepared from HeLa cells overexpressing FLAG-CerS2 and myc-CerS6 32 h after exposure to 0 or 10 Gy. FLAG-CerS2 was immunoprecipitated from lysate with anti-FLAG conjugated agarose beads and co-immunoprecipitation of myc-CerS6 determined by SDS-PAGE/western blotting with anti-myc monoclonal antibody. Similar experiments were performed with unirradiated and irradiated cells overexpressing (B) FLAG-CerS5 and myc-CerS6 and (C) HA-CerS2 and FLAG-CerS5, where FLAG-CerS5 and HA-CerS2 were immunoprecipitated using anti-FLAG and anti-HA conjugated agarose beads, respectively. Co-immunoprecipitation of myc-CerS6 and FLAG-CerS5 were determined with (B) anti-myc and (C) anti-FLAG western blots, respectively. Western blots represent 1 of 2 similar experiments, and binding was confirmed with reverse IP/western blot antibody combinations. Overexpression of tagged CerS proteins was confirmed in all lysates by direct western blot.