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. Author manuscript; available in PMC: 2009 Oct 5.
Published in final edited form as: Nat Cell Biol. 2008 Feb 10;10(3):272–282. doi: 10.1038/ncb1689

Figure 5. XTP3-B and OS-9 interact with ER quality control components.

Figure 5

a. S-tagged versions of OS-9.1 & 2, XTP3-B and EDEM1 were expressed in HEK293 cells, lysed in 1% CHAPS and affinity purified by S-protein agarose. Samples were separated by SDS-PAGE and westerns blots performed with antibodies for Hrd1, SEL1L, GRP94 and S-tag to detect coprecipitating proteins. Hairline indicates where western blot exposures were joined. Full scans are presented in Supplemental Information (Fig. S5). b. Immunprecipitations from 1% TritonX-100 detergent lysate of HEK293 cells with anti-GRP94, anti-SEL1L and a control antibody (anti-HA). Western blots were probed with antibodies for OS-9, GRP94 and SEL1L. c. Coexpression of S-tagged OS- 9.1 and OS-9.2 with indicated shRNA (described in Fig. 4b) in HEK293 cells with affinity purification as in Fig. 5a. Western blots were probed with antibodies against S-tag, GRP94, BiP and SEL1L. d. Composite data of pulse-chase assays for NHK coexpressed with a CTRL (open square) or GRP94 (closed triangle) shRNA. Data are presented as in Fig. 3c. Mean and S.E.M. were determined from 4 individual experiments.