Fig. 2

STYK1 promotes canonical Wnt/β-catenin signaling. a Heatmap of gene expression of PANC-1 cells after STYK1 depletion. b, c Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) pathway enrichment analysis of differentially expressed genes after STYK1 depletion. d, e GSEA analysis of enriched gene set in the Wnt/β-catenin signaling comparison of normal and STYK1-depleted PANC-1 cells and TCGA pancreatic cancer database with distinguishing STYK1 level. f TOP/FOPflash reporter assays quantified Wnt activity in PANC-1/AsPC-1 cells in response to Wnt3a treatment (100 ng/mL, 4 h; n = 6). g, h 7TGC plasmid schematic and fluorescence imaging of PANC-1 cells with STYK1 knockdown or WT/K147R mutant expression, with quantified GFP/RFP intensity ratios (n = 3). Scale bar: 100 μm. i, j mRNA (n = 6) and protein levels of Wnt targets (CyclinD1, C-myc, Axin2) in PANC-1 cells following STYK1 depletion with or without Wnt3a treatment. k Representative immunohistochemical images of Axin2, CyclinD1, and C-myc in excised tumors tissues. Scale bar: 100μm. l Confocal microscopy analysis of β-catenin localization in PANC-1 cells following STYK1 depletion with or without Wnt3a treatment. Scale bar: 50 μm. m Western blot analysis of nuclear β-catenin levels in STYK1-depleted PANC-1 cells with or without Wnt3a treatment. n Western blot analysis of nuclear β-catenin levels in HEK293T cells overexpressing WT or K147R mutant STYK1 with or without Wnt3a treatment. o, p Endogenous TCF4/β-catenin interaction in PANC-1 cells following STYK1 depletion or WT/K147R mutant overexpression. Data were represented as mean ± SD, *p < 0.05; **p < 0.01; ***p < 0.001