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Figure 7


Fig. 7. The absence of ubiquitylated AE1 in transfected HEK293 cells. (A,B) HEK293 cells transiently transfected with wild-type (WT) and R664X (RX) AE1, their EGFP-tagged forms (EGFP-WT and EGFP-RX), EGFP-{Delta}F508-CFTR (EGFP-{Delta}F508), or empty vectors (Mock) were incubated for 8 hours in the absence (-) or presence (+) of 10 µM lactacystin. AE1 and EGFP-{Delta}F508-CFTR were immunoprecipitated with cdb3-64 and the anti-CFTR, respectively. AE1 and EGFP-{Delta}F508-CFTR in whole-cell lysates (A) and immunoprecipitates (B) were detected by immunoblotting with cdb3-64 and anti-CFTR (A) or anti-38K and anti-GFP (B, left panel), respectively. Ubiquitylated proteins in the immunoprecipitates were also reacted with the anti-ubiquitin (anti-Ub) antibody (B, right panel). Signals corresponding to high-molecular-weight species (HMW) were seen only on blots of EGFP-{Delta}F508-CFTR. The asterisk indicates a non-specific product.





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