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. Author manuscript; available in PMC: 2018 Mar 7.
Published in final edited form as: Structure. 2011 Mar 9;19(3):337–348. doi: 10.1016/j.str.2010.12.022

Figure 3. Mutations in MESD Reduce Trafficking of Full-Length LRP6.

Figure 3

The ability of mutant MESD to traffic LRP6 was determined by assessing the maturation of LRP6 in cotransfection assays with MESD. (A) Representative western blot analyses of LRP6 maturation. The top 6% SDS-PAGE gel (>190 kDa) separates the glycosylated mature LRP6-Rho (upper band, magenta) from the lower molecular weight ER-retained LRP6-Rho (lower band, magenta). The lower 12% SDS-PAGE gel (<37 kDa) resolves hIgG (transfection control, green) and FLAG-tagged MESD (magenta). Proteins were visualized, and fluorescence was quantified using the Odyssey Infrared Imaging System (LI-COR Biosciences). (B) The efficiency of LRP6 maturation was determined by calculating the percentage of the mature membrane form (upper band) out of total LRP6 (upper and lower bands). Asterisks (*) highlight mutations that result in a significant (<0.05) reduction or enhancement of LRP6 maturation compared to wild-type (as determined by one-way ANOVA). Data and error bars indicate average ± standard deviation. Note that F141R/M151R is not significantly different compared to individual F141R or M151R mutations.