Fig 2: SOCS3 binds to PTK6 and inhibits PTK6 expression.A Network diagram of proteins related to PTK6 from the BioGRID database. B The Venn diagram of IP-MS and BioGRID. C The results of the SDS-PAGE visualized by Coomassie Brilliant Blue staining after immunoprecipitating. D, D’ Coimmunoprecipitation analysis of PTK6 and SOCS3 in UM cells. E, F and E’, F’ Western blotting analysis of SOCS3 and PTK6 in UM cells overexpressing SOCS3. G, H and G’, H’ Western blotting analysis of SOCS3 and PTK6 in UM cells with SOCS3 knockdown. The above data were analyzed using one-way ANOVA with Bonferroni. (D–H: SP6.5 cell; D’–H’: MUM-2B cell; data were presented as the mean ± SD; n = 3; *p < 0.05, **p < 0.01, ***p < 0.001).

Fig 3: PTK6 promotes the proliferation, migration, and invasion of UM cells in vitro and in vivo.A, A’ The expression of PTK6 mRNA in UM cells with PTK6 knockdown. B, C and B’, C’ Western blotting analysis of PTK6 in UM cells with PTK6 knockdown. The above datas were analyzed using one-way ANOVA with Bonferroni. D–H and D’–H’ Analysis of the proliferation, migration, and invasion of UM cells with PTK6 knockdown. D and D’ were analyzed using two-way ANOVA with Bonferroni. E–H and E’–H’ were analyzed using one-way ANOVA with Bonferroni. I, I’ The expression of PTK6 mRNA in UM cells overexpressing PTK6. J, K and J’, K’ Western blotting analysis of PTK6 in UM cells overexpressing PTK6. The above data were analyzed using one-way ANOVA with Bonferroni. L–P and L’–P’ Analysis of the proliferation, migration, and invasion of UM cells overexpressing PTK6. L and L’ were analyzed using two-way ANOVA with Bonferroni. M–P and M’–P’ were analyzed using one-way ANOVA with Bonferroni. Q’ The volume of tumors formed in the LV-NC group and LV-PTK6 group. The data were analyzed using two-way ANOVA with Bonferroni. R’ Photographic images of tumors from the LV-NC group and LV-PTK6 group. S’ The weight of tumors formed in the LV-NC group and LV-PTK6 group. The data were analyzed using Student’s t-test. (A-P: SP6.5 cell; A’–S’: MUM-2B cell; scale bar: 100 µm; Data were presented as the mean ± SD; n = 3–6; *p < 0.05, **p < 0.01, ***p < 0.001).

Fig 4: Proposed schematic illustration of the PTK6 mechanism: PTK6 inhibits autophagy to promote UM tumorigenesis by binding to SOCS3 and regulating mTOR phosphorylation.

Fig 5: PTK6 promotes the proliferation, migration, and invasion of UM cells by inhibiting autophagy.A, B and A’, B’ Western blotting analysis of LC3, PTK6, and P62 in the Si-PTK6 groups and Si-NC group after treatment with DMSO or 3-MA. The data were analyzed using two-way ANOVA with Bonferroni. C–G and C’–G’ Proliferation, migration, and invasion analyses of the Si-PTK6 groups and Si-NC group after treatment with DMSO or 3-MA. The data were analyzed using two-way ANOVA with Bonferroni. H, I and H’, I’ Western blotting analysis of LC3, PTK6, and P62 in the LV-NC group and LV-PTK6 group after treatment with DMSO or rapamycin. The data were analyzed using two-way ANOVA with Bonferroni. J–N and J’–N’ Proliferation, migration, and invasion analyses of the LV-NC group and LV-PTK6 group after treatment with DMSO or rapamycin. The data were analyzed using two-way ANOVA with Bonferroni. A–N: SP6.5 cell; A’–N’: MUM-2B cell; scale bar: 100 µm; data were presented as the mean ± SD; n = 3; ns no significant difference, *p < 0.05, **p < 0.01, ***p < 0.001).