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. 2006 Oct 6;103(42):15657–15662. doi: 10.1073/pnas.0604621103

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© 2006 by The National Academy of Sciences of the USA

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Fig. 3. — The ICDI domain binds to the EF hand motif of Cav1.4. (A) Scheme of the Cav1.4 C terminus starting after the IVS6 segment. (B Upper) Co-IP of HEK293 cells coexpressing triple flag-tagged ICDI together with the myc-tagged CT1610Stop or CT1610Stop,ΔEF. Lysates were immunoprecipitated with anti-myc (lanes 4 and 5) or anti-ras (control, lanes 6 and 7), blotted and probed with anti-flag. Lane 8: negative control. (B Lower) Immunoprecipitation of HEK293 cells expressing ICDI and myc-tagged peptides C or A. Co-IP has been carried out as in Upper. (C) ICDI interaction assay with GST-tagged fragments of the proximal C terminus. GST fusion proteins were incubated with bacterial lysates containing flag-tagged ICDI. Immunoblotting was performed with an anti-flag antibody. In the last lane, pure ICDI was blotted to demonstrate specificity of the assay. Each experiment was repeated at least three times.