Fig. 5.

Pharmacological inhibition of TGF-β signaling blocks LAMC2-induced metastasis. a Volume of tumors formed following subcutaneously injection of EGFP⁺ and EGFP⁻ cells in nude athymic mice. n ≥ 10. b Representative H&E-stained sections of xenografts derived from EGFP⁺ or EGFP⁻ cells. c qPCR analysis for LAMC2, EMT, MMP2 and MMP10 gene expression in EGFP⁺ or EGFP⁻ cells, isolated from tumors. Data are normalized to GAPDH and are presented as fold change relative to EGFP⁻. d qPCR analysis or CD44 and CD133 expression in EGFP⁺ or EGFP⁻ cells isolated from tumors. Data are normalized to GAPDH and are presented as fold change in gene expression relative to EGFP⁻. e Number of tumors generated by subcutaneous injection of EGFP⁺ or EGFP⁻ cells. f Enrichment plot for EGFP⁺ versus EGFP⁻ cells isolated by FACS from subcutaneous tumors. g Representative immunofluorescence images for pSMAD2 (violet), LAMC2 (green) and nuclei (blue, DAPI) in tumor sections derived from EGFP⁻ or EGFP⁺ cells subcutaneously xenografted in nude athymic mice. h qPCR analysis for LAMC2 expression in PDAC cells treated with 10 ng/ml of rTGF-β1 in the presence or absence of 80 μM Vactosertib. Data are normalized to GAPDH and are presented as fold change in gene expression relative to control. i Representative H&E-stained sections of lungs following tail vein injection of EGFP⁺ or EGFP⁻ tumor cells. Mice were treated with Vactosertib (40 mg/kg mice) or vehicle. *p < 0.05, **p < 0.005, ***p < 0.0005. n $\ge 5.$ Statistical significance was assessed by Student's t-test