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. Author manuscript; available in PMC: 2015 Apr 15.
Published in final edited form as: Biochem J. 2014 Apr 15;459(2):417–425. doi: 10.1042/BJ20131037

Figure 4. Contribution of protein acyl transferases to CFTR biogenesis.

Figure 4

Western blot of HEK293 cells co-transfected with HA-tagged DHHC proteins and either wild-type (A) or F508del (B) CFTR indicate an increase in band B following overexpression of DHHC-7. Co-immunoprecipitation (DHHC proteins used as anchor) identified a robust interaction between DHHC-7 and wild-type CFTR (C) as well as F508del CFTR (D). Metabolic labeling with 3H-palmitic acid in the presence of DHHC-7 resulted in enhanced CFTR palmitoylation (E), indicated by labeling after a 3–4 week exposure as compared to the 3–4 months required for CFTR labeling without DHHC-7. To verify palmitoylation, post-IP samples were split and treated with 1 M Tris-Cl (control) or 1 M hydroxylamine (pH 7.4) to remove palmitate via cleavage of the thioester bond.