Fig 2: Ectopic expression of COL5A2 promotes cancer cell proliferation and upregulates WNT/ß-catenin and PI3K/mTOR pathway. (A) COL5A2 was ectopically expressed in HEK293T and colon cancer cell lines SW620 and HCT116 and the cell variability was measured using MTT method in different days. (B and C) Ectopic expression of COL5A2 and selected signaling molecules were measured by Western blot. (D) Western blot analyses of DDR1 and pY792 DDR1 in COL5A2 overexpressing CRC cells. The Western blot densitometry was calculated using Multi Gauge V3.0 and the fold change was normalized using the blank vector transfection. COL5A2 protein was probed using two antibodies. Active ß-catenin was the endogenous ß-catenin protein when residues Ser33, Ser37, and Thr41 are not phosphorylated. The arrow heads indicate the positions of GSK3ß.

Fig 3: The silencing of TM4SF1 induces cell cycle G2/M arrest, apoptosis and affects DDR1 signaling and apoptotic markers in NSCLC cell lines. a Representative distribution of A549 and H1299 cell lines both in NC and TM4SF1 silencing cells (left); The distribution and percentage of cells in the G1, S and G2/M phases of the cell cycle are indicated (right).b Knocking down of TM4SF1 sensitized NSCLC cells to cisplatin and paclitaxel, The cell viability was detected by MTS assay after cells were treated with chemotherapy drugs paclitaxel for 24 h or cisplatin for 12 h. All experiments were performed in triplicate, respectively. *p < 0.05, **p < 0.01, ***p < 0.001. c Induction of apoptosis detected by flow cytometric analysis with Annexin V-FITC and PI-staining (right). Quantitative analysis of apoptosis (**p < 0.01, ***p < 0.001). d Expression levels of Casp7, 9, 3, cleaved-PARP were evaluated by western blot in NC and TM4SF1 silencing cells. And the impact of silencing TM4SF1 on the expression of DDR1 and the phosphorylation of its downstream targets AKT, ERK and mTOR. e Inhibitor down-regulated DDR1, mTOR and AKT and Knocking down TM4SF1 can further lower the expression of this pathway after inhibitor treatment

Fig 4: Effects of TFAP2A-mediated DDR1 regulation on collagen fiber arrangement and immune cell infiltration in breast cancer. Note: (A) RT-qPCR detected the expression levels of TFAP2A and DDR1 in each group of EO771 cells; (B) Masson staining detected the collagen fiber arrangement in each group of BC-MPS treated with different EO771 cells (×100); (C) Tumor volume of each group of breast cancer mice; (D) Tumor weight of each group of breast cancer mice; (E) Photos of tumors taken from each group of mice; (F) RT-qPCR detected the expression levels of TFAP2A and DDR1 in each group of breast cancer mice tumor tissues; (G) Immunohistochemistry detected the content of Collagen IV in ECM of each group of breast cancer mice tumor tissues (×200); (H) Masson staining detected the collagen fiber arrangement in each group of breast cancer mice tumor tissues (×400); (I) Immunohistochemistry detected the number of CD8-positive cells in each group of breast cancer mice tumor tissues (×200); (J) CD8 + T cell migration experiment detected the migration of CD8 + T cells in each group of breast cancer mice tumor tissues after decellularized ECM treatment. * indicates P < 0.05 compared with the control group. All cell experiments were repeated 3 times; each group consisted of 5 mice

Fig 5: Regulation of DDR1 by transcription factor TFAP2A. Note: (A) Correlation analysis of DDR1 and transcription factors MAZ, FOXA1, ZBTB7B, TFAP2A, GATA3, and ZNF263 in breast cancer-related TCGA dataset, including 1096 breast cancer tissues; (B) RT-qPCR detected the expression levels of MAZ, FOXA1, ZBTB7B, TFAP2A, GATA3, and ZNF263 in breast cancer cells (EO771, EMT6, and 4T1.2); (C) RT-qPCR detected the expression levels of TFAP2A in each group of EO771 cells after silencing or overexpression; (D) RT-qPCR detected the expression levels of DDR1 in each group of EO771 cells after silencing or overexpression; (E) Dual-luciferase assay detected the activity of DDR1 promoter in each group of EO771 cells after silencing or overexpression of TFAP2A; (F) ChIP assay detected the enrichment of TFAP2A on DDR1 promoter in each group of EO771 cells after silencing or overexpression of TFAP2A. * indicates P < 0.05, ** indicates P < 0.01 compared with the control group. All cell experiments were repeated 3 times