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. 2013 Mar 15;24(6):715–733. doi: 10.1091/mbc.E12-07-0537

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© 2013 Johnson et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 7: — Detergent solubility of knocked-in WT-Cx43 and its sorting motif and serine mutants. (A) CP43KD cells expressing knocked-in WT-Cx43 and various mutants were extracted in situ with 1% TX100 and immunostained for Cx43. Note that only knocked-in YA/VD-Cx43 (YA/VD) assembles into detergent-resistant gap junction puncta. Note also that intracellular puncta of knocked-in WT-Cx43 (WT) and all other mutants are lost upon extraction with 1% TX100. (B) Western blot analysis of Cx43 in Total (T), TX100-soluble (S), and TX100-insoluble (I) fractions from CP43KD cells expressing knocked-in WT-Cx43 and the mutants. Note that only cells expressing knocked-in YA/VD-Cx43 have a discernible detergent-insoluble fraction. The blots were stripped and reprobed for β-actin to verify equal loading.