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. Author manuscript; available in PMC: 2011 May 1.
Published in final edited form as: Channels (Austin). 2010 May 1;4(3):192–202. doi: 10.4161/chan.4.3.11867

Figure 1. Rem interacts with the proximal and distal domains of CCT.

Figure 1

A, Schematic of the CCT truncation mutants with the Rem interaction status indicated on the right. B, TsA201 cells were transiently co-transfected with expression vectors encoding 3xFlag-tagged Rem and either pCDNA3.1+3xHAa (empty vector), HA-CCT-FL or the indicated HA-tagged CCT truncation mutants. 48 h post-transfection, cells were harvested, and cell lysate (0.5 mg) was subjected to immunoprecipitation with anti-HA antibody as described under “Materials and Methods”. The entire bound fraction or a portion of the unbound fraction (2.5 µl) was analyzed by immunoblotting with biotin-Flag to detect Rem. C, Cell lysate (5 µg) was immunoblotted with anti-HA antibody to monitor expression of CCT-FL and the corresponding truncation mutants used in panel B. D, TsA201 cells were transiently co-transfected with vectors expressing Flag-Rem and either pCDNA3.1+3xHAa, HA-Cav1.2 CCT or HA-Cav3.2 CCT. Co-immunoprecipitation was performed using HA antibody as described in B and Rem binding examined by biotin-Flag immunoblotting. Results in each panel are representative of three independent experiments.