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. 2023 Sep 27;11:1236356. doi: 10.3389/fcell.2023.1236356

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Copyright © 2023 Singh, Walter, Elcheva, Imamura Kawasawa and Spiegelman.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Global role of IGF2BP1 in Wnt/β-catenin-regulated genes in CRC cells. (A) Pictorial diagram illustrating the experimental setup: DLD1 cells were transformed and treated with doxycycline to induce the expression of dnTcf7L2 and GFP in DLD1-dnTCF7L2-GFP cells, and dnTcf7L2 and Igf2bp1 in DLD1-dnTCF7L2-IGF2BP1 cells. (B) Immunoblot assay showing the expression levels of Tcf7L2 and Igf2bp1 after doxycycline treatment. β-actin levels were used as a loading control. (C) TOP/FOP-Flash assay indicating the extent of Wnt signaling inhibition upon induction of dnTcf7L2 and GFP in DLD1-dnTCF7L2-GFP cells, and dnTcf7L2 and Igf2bp1 in DLD1-dnTCF7L2-IGF2BP1 cells. Data are presented as means ± SD (n = 3). (D) Pie chart displaying differentially expressed genes upon Wnt signaling inhibition in DLD1-dnTCF7L2-GFP cells. (E) Bar plot illustrating IGF2BP1-dependent/independent genes in DLD1-dnTCF7L2-IGF2BP1 cells that were upregulated in DLD1-dnTCF7L2-GFP. (F) Bar plot depicting IGF2BP1-dependent/independent genes in DLD1-dnTCF7L2-IGF2BP1 cells that were downregulated in DLD1-dnTCF7L2-GFP. (G) Pie chart showing the total number of IGF2BP1-dependent and independent genes regulated by Wnt signaling. (H) Gene ontology analysis of subgroup of 177 downregulated genes (F) regulated by Wnt signaling and controlled by IGF2BP1.(I) Proliferation measured as % confluence of DLD1, DLD1-dnTCF7L2, and DLD1-dnTCF7L2-IGF2BP1 using the Incucyte Live-Cell Analysis System. (J) Venn diagram illustrating the overlapping genes between 177 Wnt target genes (F) regulated by IGF2BP1 and TCF7L2-ChIP-seq genes (reported in a published article).