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. Author manuscript; available in PMC: 2018 Mar 1.
Published in final edited form as: Mitochondrion. 2016 Aug 31;33:84–101. doi: 10.1016/j.mito.2016.08.017

Figure 2. BKCa interacts with Tom22 in HEK293T cells.

Figure 2

Cells were co-transfected with BKCa, BKCa-DEC and Tom22 in different combinations (+, present; -, absent). Co-immunoprecipitated products are highlighted with squares. (A) Immunoblot demonstrating that BKCa and BKCa-DEC pull down Tom22. BKCa constructs were immunoprecipitated with anti-HA pAb and detected with anti-HA mAb (arrows) and co-immunoprecipitated Tom22 was detected with anti-c-Myc mAb (arrowhead, squares). Primary antibodies were incubated at the same time. (B) Immunoblot showing proper expression of BKCa constructs (arrows) and Tom22 (arrowhead) in the input lysates. (C) BKCa and BKCa-DEC efficiency in pulling down Tom22 is similar (n=3 each). Co-IP efficiency was determined by first subtracting background signals, which in this case were negligible, and calculating the ratio of the band intensities of co-immunoprecipitated Tom22 (i.e. A, squares) to corresponding immunoprecipitated BKCa constructs (i.e.. A, arrows), and normalized to Tom22 intensity in the corresponding input lysates (i.e. B, arrowhead). In this and following figures, image analysis was performed with MetaMorph Image Analysis Software (Molecular Devices).