Fig 2: The NF2 LOF mutations induce an oncogenic role of NF2 through the VANGL-JNK pathway.a Illustration of NF2 domains and its identified LOF missense mutations. b The NF2 LOF mutants fail to rescue YAP cytoplasmic localization in the NF2 KO cells. Scale bar, 30 μm. Arrows showed the cells expressing the indicated constructs. c Reconstituting the NF2 LOF mutants fail to rescue YAP S127 phosphorylation in the NF2 KO cells. A representative blot of two independent experiments is shown. d, e The NF2 LOF mutations inhibit its interaction with LATS1 and AMOT. HEK293T cells were transfected with the indicated HA-NF2 constructs and SFB-tagged LATS1 (d) or AMOT (e) and subjected to pulldown assay. A representative blot of two independent experiments is shown. f NF2 LOF mutants induces the phosphorylation of JNK and c-Jun in MCF10A cells. A representative blot of two independent experiments is shown. g, h The NF2 LOF mutants promote MCF10A acini growth. The representative images were shown (g). The acini size was measured and quantified (mean ± s.d., n = 40 acini per group) (h). ***p < 0.001 (two-tailed Student’s t-test). Scale bar, 400 μm. i, j The NF2 LOF mutations enhance its interaction with VANGL1. The interaction between the indicated HCIPs and NF2 LOF mutants was normalized and shown as a heatmap (i). HEK293T cells were transfected with SFB-VANGL1 and the indicated HA-NF2 LOF mutants and subjected to pulldown assay (j). A representative blot of two independent experiments is shown. k The NF2 LOF mutants activate JNK through VANGL1. The MCF10A cells stably expressing NF2 and its LOF mutants were transduced with control and VANGL1 shRNAs. A representative blot of two independent experiments is shown. l The MCF10A cells stably expressing the NF2 LOF mutants were treated with JNK-IN-8 (10 μM) for 4 h and subjected to Western blot. A representative blot of two independent experiments is shown. m, n VANGL1 and JNK are required for the NF2 LOF mutants-induced MCF10A acini growth in Matrigel. The representative images were shown (m). The acini size was measured and quantified (mean ± s.d., n = 40 acini per group) (n). ***p < 0.001 (two-tailed Student’s t-test). Scale bar, 400 μm. o The NF2 LOF mutants activate JNK in IOMM-Lee cells. A representative blot of two independent experiments is shown. p, q Treatment with JNK-IN-8 inhibits NF2 LOF mutants-induced IOMM-Lee xenograft tumor growth. The collected tumors treated with DMSO or JNK-IN-8 (10 mg/kg) were shown (p) and tumor weight was measured and quantified (mean ± s.d., n = 6 mice per group) (q). ns, no significance. *p < 0.05, **p < 0.01, ***p < 0.001 (two-tailed Student’s t-test). Source data are provided as a Source Data file.