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. 2024 Jun 26;115(9):3013–3025. doi: 10.1111/cas.16258

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© 2024 The Author(s). Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.

This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.

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The RECK/GPR124 complex mediates WNT signaling. (A) RECK and Myc‐tagged GPR124 simultaneously expressed in 293T cells and immunoprecipitated (IP) by antibodies indicated. The precipitants were then immunoblotted (IB) by antibodies indicted (left). The expression of proteins in the input whole‐cell lysates (right). (B) TOP/FOP canonical WNT/β‐catenin reporter assay of 293T cells transduced with pCXN2‐RECK (N = 3). (C) TOP/FOP assay of 293T cells transduced with Myc‐tagged GPR124 in 293T cells (N = 3). (D) TOP/FOP assay of 293T cells transduced with pCXN2‐RECK and pcDNA3.1 Myc‐GPR124 mixed at the indicated ratio (N = 3). (E) TOP/FOP assay of 293T cells transduced with pCXN2‐NEO and pcDNA3.1 mixed at the indicated ratio (N = 3). (F) TOP/FOP assay of 293T cells transduced with pCXN2‐RECK, RECK mutants and pcDNA3.1 Myc‐GPR124 mixed at the ratio of 1:9 and treated with or without 10 ng/mL of WNT7A (N = 3). Control columns appear twice for the convenience of assessing significance between control and N200Q and M207W columns. (G) RECK (WT and mutants) and Myc‐tagged GPR124 were simultaneously expressed in 293T cells and immunoprecipitated (IP) by the antibodies indicated. The precipitants were then immunoblotted (IB) by the antibodies indicated (upper). The expression of proteins in the input whole‐cell lysates were detected by the corresponding antibodies (lower). * p < 0.05 against vector control; N.S., not significant.