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. Author manuscript; available in PMC: 2017 Oct 18.
Published in final edited form as: Cell Rep. 2016 Nov 1;17(6):1546–1559. doi: 10.1016/j.celrep.2016.10.037

Figure 2. Mammalian SEIPIN and GPAT3/4 Physically Interact.

Figure 2

(A) Co-immunoprecipitation by FLAG and hemagglutinin (HA) antibody of overexpressed FLAG-tagged GPAT3 or GPAT4 and HA-tagged SEIPIN, respectively, from transfected MEF cell lysates.

(B and C) Immunoblotting for overexpressed HA-tagged mutant or wild-type SEIPIN and FLAG-tagged GPAT3 and GPAT4, respectively, after HA-immunoprecipitation from 3T3-L1 cells. See also Figure S2.

(D) Endogenous GPAT4 and GPAT3 proteins co-immunoprecipitate with overexpressed SEIPIN from 3T3-L1 adipocytes stably transfected with HA- or FLAG-SEIPIN.

(E) 3T3-L1 adipocytes expressing FLAG-SEIPIN, FLAG-tagged biotin ligase BirA*, or FLAG-tagged BirA*-SEIPIN were treated with 50 μM biotin for 24 hr. The biotinylated proteins were pulled down using streptavidin-conjugated agarose from biological duplicates. Elutes were subjected to western blot analysis. All experiments were performed in triplicate.