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. 2021 Mar 2;220(4):e202012112. doi: 10.1083/jcb.202012112

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© 2021 Nong et al.

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Figure 4. — The LLPS property of Axin is needed to inhibit Wnt signaling. (A) Axin1 KO HEK293T cells were transfected with TopFlash-luciferase reporter and WT Axin1-EGFP or its mutants, then treated with or without Wnt3a conditional medium for 12 h and harvested for luciferase determination. Statistical analyses were performed with two-tailed unpaired t test. Data are shown as mean ± SD (n = 3). The stars (*) indicate the significant differences between each mutant group with the WT Axin1 group, while the hash symbols (#) indicate the significant differences between AD7^TDP43 or AD7^hnRNPA1 with the AD7 group. **, P < 0.01; ***, P < 0.001; ###, P < 0.0001. (B) Axin1 KO HEK293T cells transfected with WT Axin1-EGFP or its mutants were treated with 2.5 µg/ml cycloheximide (CHX) for the indicated time before harvesting for immunoblotting. (C) The Xenopus embryos were dejellied 30 min after fertilization and injected with mRNA encoding WT Axin1 or mutants. Embryos were cultured to the tailbud stage (stage 32) for phenotyping. Embryos with different morphologies and their percentages are shown. The histogram summarizes results from two independent experiments. β-cat, β-catenin; Vec, vector; Rel., relative.