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. Author manuscript; available in PMC: 2019 Feb 1.
Published in final edited form as: Biochim Biophys Acta Mol Cell Res. 2018 Nov 14;1866(2):252–263. doi: 10.1016/j.bbamcr.2018.11.005

Fig. 3.

Fig. 3

Verification of ATB0,+ interaction with SEC24 proteins. (A) HEK293 cells stably transfected with p3xFLAG-CMV14/B0,+ vector were treated either with DMSO alone or with 200 nM PMA (PKC activator). ATB0,+ has been immunoprecipitated with anti-FLAG antibody immobilized on agarose resin, as described in Materials and methods and eluted with FLAG peptide. The eluates (IP) were analyzed with antibodies against SEC24 isoforms (Cell Signaling Technology). (B) Analysis of SEC24C co-precipitating with ATB0,+ (Novus Biologicals anti-SEC24C antibody). For better separation of SEC24C double bands, the lower acrylamide concentration (8%) was used in the separating gel. Cells were either transfected with p3xFLAG-CMV14 vector (V) or with p3xFLAG-CMV14/B0,+ vector (B0,+). T, corresponds to Western blot analysis of the total cell extract, IP for the analysis of fraction eluted with FLAG peptide. As controls, immunoprecipitates were analyzed for the presence of ATB0,+ with anti-FLAG antibodies.