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. Author manuscript; available in PMC: 2020 Mar 1.
Published in final edited form as: Biochim Biophys Acta Mol Cell Biol Lipids. 2018 Dec 6;1864(3):245–259. doi: 10.1016/j.bbalip.2018.11.007

Figure 5. The ORM proteins mediate oligomerization of SPT.

Figure 5.

A. Yeast Lcb1-GFP was co-expressed with either Lcb1-FLAG (Panels 1-3) or Lcb1ΔTMD1-FLAG (Panel 4) in wild-type (Panels 1 and 4), orm1Δorm2Δ (Panel 2), or Sac1Δ (Panel 3) mutant cells. The ability of Lcb1-FLAG or Lcb1ΔTMD1-FLAG to coimmunoprecipitate Lcb1-GFP was assessed by immunoblotting after adsorption of solubilized microsomal proteins to anti-FLAG beads. B. The ability of Lcb1-GFP or Lcb1ΔTMD1-GFP to coimmunoprecipitate Lcb1-FLAG was determined by adsorption of solubilized microsomal proteins from Panel 1 or Panel 4 in A. to anti-GFP beads followed by SDS-PAGE and immunoblotting with anti-FLAG antibodies. C. Yeast microsomes were prepared from lcb1Δsac1Δ mutant cells expressing Lcb1- FLAG and HA-Sac1, solubilized with 2% DIG, 1% DDM, 0.1% SML or 1% Triton X-100 (TX100) at 4 °C and incubated with anti-FLAG beads. Adsorbed proteins were resolved by SDS-PAGE and visualized by immunoblotting with antibodies to the proteins indicated on the right side of the panel.