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. 2019 Dec 27;219(2):e201903017. doi: 10.1083/jcb.201903017

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© 2019 Mis et al.

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Figure 5. — IPO11 is required for CRC organoid growth. (A) Crystal violet stain of colony formation of DLD-1 βcatenin-DEADPOOL cells transduced with LacZ, IPO11 #1, and IPO11 #2 gRNA. (B) RT-qPCR of ASCL2 in CSC171C organoid line transduced with LacZ, IPO11 #1, or IPO11 #2 gRNA. Mean fold change ± SD, representative of n = 3 independent experiments. Statistical analysis was performed by one-way ANOVA Dunnett’s test. *, P ≤ 0.05; **, P ≤ 0.01. (C) Fluorescence images of CSC171C organoid line expressing Cas9 and gRNA for LacZ, CTNNB1 #1, IPO11 #1, or IPO11 #2. (D) Points represent mean fold change ± SD of fluorescence of C representative of n = 3 independent experiments. Statistical analysis was performed by one-way ANOVA Dunnett’s test. ***, P ≤ 0.001; ****, P ≤ 0.0001. (E) Proposed model of βcatenin nuclear translocation through direct binding of βcatenin armadillo repeats and nuclear pore proteins via a Ran-independent mechanism. IPO11 directly binds βcatenin in the cytoplasm to mediate βcatenin nuclear import. In the nucleus, IPO11 binds to Ran-GTP, which leads to the dissociation of the cargo–importin-β complex. βcatenin accumulates in the nucleus.