Fig 2: Overexpressing SOCS3 can partially attenuate the PTK6-induced promotion of UM cell proliferation, migration, and invasion in vitro and in vivo.A, B and A’, B’ Western blotting analysis of SOCS3 and PTK6 in the LV-NC group or LV-PTK6 group supplemented with LV-NC and LV-SOCS3. C–G and C’–G’ Proliferation, migration, and invasion analysis of the LV-NC group or LV-PTK6 group treated with LV-NC and LV-SOCS3. The above data were analyzed using two-way ANOVA with Bonferroni. H’ The volume of tumors formed in the LV-PTK6/LV-NC group and LV-PTK6/LV-SOCS3 group. The data were analyzed using two-way ANOVA with Bonferroni. I’ Photographs of tumors from the LV-PTK6/LV-NC group and LV-PTK6/LV-SOCS3 group. J’ The weight of tumors formed in the LV-PTK6/LV-NC group and LV-PTK6/LV-SOCS3 group. The data were analyzed using Student’s t-test. (A–G: SP6.5 cell; A’–J’: MUM-2B cell; scale bar: 100 µm; data were presented as the mean ± SD; n = 3–6; ns no significant difference, *p < 0.05, **p < 0.01, ***p < 0.001).

Fig 3: PTK6 promotes mTOR phosphorylation.A, B and A’, B’ Western blotting analysis of mTOR and p-mTOR in UM cells with PTK6 knockdown. C, D and C’, D’ Western blotting analysis of mTOR and p-mTOR in UM cells overexpressing PTK6. The above data were analyzed using one-way ANOVA with Bonferroni. E, F and E’, F’ Western blotting analysis of mTOR and p-mTOR in the four cell lines: LV-NC/LV-NC, LV-PTK6/LV-NC, LV-NC/LV-SOCS3, and LV-PTK6/LV-SOCS3. The data were analyzed using two-way ANOVA with Bonferroni. (A–F: SP6.5 cell; A’–F’: MUM-2B cell; data were presented as the mean ± SD; n = 3; ns no significant difference, *p < 0.05, **p < 0.01, ***p < 0.001).

Fig 4: PTK6 inhibits autophagy in UM cells.A, B and A’, B’ Western blotting analysis of LC3 and P62 in UM cells with PTK6 knockdown. C, D and C’, D’ Immunofluorescence analysis of autophagosomes in UM cells with PTK6 knockdown. The above data were analyzed using one-way ANOVA with Bonferroni. E, F and E’, F’ Western blotting analysis of LC3 and P62 in UM cells overexpressing PTK6. G, H and G’, H’ Immunofluorescence analysis of autophagosomes in UM cells overexpressing PTK6. The above data were analyzed using one-way ANOVA with Bonferroni. (A-H: SP6.5 cell; A’–H’: MUM-2B cell; scale bar: 20 µm; Data were presented as the mean ± SD; n = 3; *p < 0.05, **p < 0.01, ***p < 0.001).

Fig 5: 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).