Fig. 2: PSPC1 or PSPC1-Y523F releases PTK6 to synergize the oncogenic effects. | Nature Communications

Fig. 2: PSPC1 or PSPC1-Y523F releases PTK6 to synergize the oncogenic effects.

From: PSPC1-interchanged interactions with PTK6 and β-catenin synergize oncogenic subcellular translocations and tumor progression

Fig. 2: PSPC1 or PSPC1-Y523F releases PTK6 to synergize the oncogenic effects.

a The PSPC1-Y523F but not PSPC1-Y383F mutation abolished the PSPC1/PTK6 interaction demonstrated by IP/Western blotting analysis. (Top) A cartoon illustrates domain structures and tyrosine residues Y383 (Red) and Y523 (Blue) in the C-terminal PSPC1 protein. b, c PTK6 suppressed the PSPC1-enhanced cell migration (b), invasion (c), in SK-hep1 cells. Mutation of PSPC1-Y523F but not Y383F abolished the PSPC1-enhanced cell migration and invasion. Data are represented as mean ± SEM (n = 4). d, e SNU-387 expressed a higher level of PTK6 could to some extent reduce PSPC1 and the PSPC1-Y383F-enhanced cell migration (d), invasion (e), but not that of PSPC1-Y523F (loss of PTK6 interaction ability). Data are represented as mean ± SEM (n = 4). f Immunofluorescence for the detection of subcellular localizations of PSPC1, p-PTK6, and PTK6 in SK-Hep1. Colors for p-PTK6 as well as PTK6, PSPC1, and DAPI are red, green, and blue, respectively. The scale bar represents 10 µm. g Western blotting analysis of EMT and cancer stemness markers in SK-hep1 cells expressing PSPC1, PSPC1-Y523F, and PTK6. h Western blotting analysis of EMT and cancer stemness markers in SNU-387 cells expressing PSPC1, PSPC1-Y523F. i Spheroids formation in different PSPC1/PTK6 SK-hep1 transfectants including phase images (Top) and graphic quantifications after two passages of spheroids (bottom). Data are represented as mean ± SEM (n = 6). j SNU387 cells expressing higher level of PTK6 could reduce PSPC1-mediated spheroids formation, but cells expressing PSPC1-Y523F could not. Data are represented as mean ± SEM (n = 6). PTK6 suppressed PSPC1-enhanced cell spheroids formation in SK-hep1 cells. The PSPC1-Y523F mutation but not Y383F potentiated spheroids formation with series dilutions for 20 days cultures. Co-expression of PSPC1-Y523F and PTK6 in SK-hep1 cells synergized oncogenic spheroid formation. All data statistics based on *p < 0.05, **p < 0.01 ***p < 0.001 calculated by one-way ANOVA with Brown–Forsythe test. Source data are provided as a Source Data file.

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