Additional file 7: Figure S7. Generation of LAMC2-EGFP knock-in human PDAC cells. (a) Flow cytometry for IRFP in PDAC cells 48 hours post-nucleofection. All cytometry gates were established based on isotype controls. (b) Flow cytometry for EGFP in PDAC cells 20 days post-nucleofection. All cytometry gates were established based on isotype controls. (c) PCR gDNA specific integration analysis. The positions of primers are indicated by arrows. (d) FACS profiles showing the expression of EGFP in...

Additional file 8: Figure S8. Characterization of human LAMC2-EGFP PDAC cells in vitro and in vivo. (a) Representative images of EGFP+ and EGFP-cells grown in Matrigel. (b) Organoid formation capacity of EGFP+ and EGFP-cells. (c) Quantification of organoid size of EGFP+ and EGFP-cells. (d) qPCR analysis for LAMC2, EMT, MMP2 and MMP10 gene expression in EGFP+ and EGFP-cells. Data are normalized to GAPDH and are presented as fold change in gene expression relative to the...

Additional file 3. Search strategies

Additional file 5. Database structure in Open Science Framework

Additional file 2. Classification of digital health technologies (DHTs)

Additional file 4. Codebook

Additional file 4. Codebook

Additional file 3: Figure S3. LAMC2 expression correlates with CSC content and function. (a) qPCR analysis of LAMC2 gene expression in adherent cells versus spheres. Data are normalized to GAPDH and are presented as fold change in gene expression relative to adherent cells (indicated as Adh). (b) Western blot analysis of LAMC2 in adherent cells versus spheres. Parallel β-ACTIN immunoblotting was performed. (c) qPCR analysis for CD44, CD133 and L1CAM genes in adherent cells versus...

Additional file 9: Figure S9. Global gene expression profiles of LAMC2-EGFP+ and EGFP--derived tumors. Heat map of differentially expressed genes in EGFP+ and EGFP- cells isolated from tumors.

Additional file 2. Classification of digital health technologies (DHTs)

Additional file 3. Search strategies

Additional file 5. Database structure in Open Science Framework

Additional file 10: Figure S10. Inhibition of transforming growth factor beta (TGF-β) signaling blocks LAMC2-induced metastasis. (a) Enrichment plots for pancreatic cancer, focal adhesion, hypoxia, MAPK signaling, glycolysis and gluconeogenesis pathways in EGFP+ versus EGFP- cells isolated by FACS from subcutaneous tumors. (b) Western blot analysis for pSMAD2 and SMAD2 in EGFP- versus EGFP+ PANC-1 and #253 cells. Parallel GAPDH immunoblotting was performed. (c) Flow cytometry quantification of EGFP and ALK5-TGFβR1 in EGFP- and EGFP+...

Additional file 4: Figure S4. Knockdown of LAMC2 does not affect cell growth. (a) Western blot analysis of LAMC2 in sh empty and LAMC2 knockdown cells. Parallel β-ACTIN immunoblotting was performed. (b) qPCR analysis for LAMC2 and CD44 gene expression in sh empty and LAMC2 knockdown cells. Data are normalized to GAPDH and are presented as fold change in gene expression relative to sh empty. (c) Representative images of sh empty and LAMC2 knockdown cells...

Additional file 6: Figure S6. Knockdown of LAMC2 affects tumorigenicity. (a) Representative images from a gelatin degradation assay of sh empty and LAMC2 knockdown cells. Nuclei were stained with Hoechst 33342 (blue), green represents actin (Alexa Fluor™ 488 Phalloidin) and red illustrates gelatin (Rodhamine). The white dashed line circles indicate the areas of degradation. (b) Degradation potential of sh empty and LAMC2 knockdown cells. (c) qPCR analysis for EMT genes in sh empty and LAMC2...

Additional file 10: Figure S10. Inhibition of transforming growth factor beta (TGF-β) signaling blocks LAMC2-induced metastasis. (a) Enrichment plots for pancreatic cancer, focal adhesion, hypoxia, MAPK signaling, glycolysis and gluconeogenesis pathways in EGFP+ versus EGFP- cells isolated by FACS from subcutaneous tumors. (b) Western blot analysis for pSMAD2 and SMAD2 in EGFP- versus EGFP+ PANC-1 and #253 cells. Parallel GAPDH immunoblotting was performed. (c) Flow cytometry quantification of EGFP and ALK5-TGFβR1 in EGFP- and EGFP+...

Additional file 1. Chronic health conditions

Additional file 8: Figure S8. Characterization of human LAMC2-EGFP PDAC cells in vitro and in vivo. (a) Representative images of EGFP+ and EGFP-cells grown in Matrigel. (b) Organoid formation capacity of EGFP+ and EGFP-cells. (c) Quantification of organoid size of EGFP+ and EGFP-cells. (d) qPCR analysis for LAMC2, EMT, MMP2 and MMP10 gene expression in EGFP+ and EGFP-cells. Data are normalized to GAPDH and are presented as fold change in gene expression relative to the...

Additional file 2: Figure S2. Increased LAMC2 expression is associated with aggressive signature. (a) Dimensional reduction plot (DimPlot) of multiple cell types identified in PDAC primary tumors by single-cell RNA sequencing (scRNA-Seq). The clusters are color-coded based on cell types identified using known cell type-specific markers and are visualized using t-Distributed Stochastic Neighbor embedding (t-SNE). (b) Violin plot showing expression levels of LAMC2 in stromal versus ductal (tumor) cells identified in PDAC primary tumors by...

Additional file 5: Figure S5. Loss of LAMC2 reduces stemness. (a) Flow cytometry for apoptotic cells as determined by AnnexinV/PI staining in control and LAMC2 knockdown cells. (b) Flow cytometry quantification of CD44 in sh empty and LAMC2 knockdown cells. (c) qPCR analysis of CD133 in sh empty and LAMC2 knockdown cells. Data are normalized to GAPDH and are presented as fold change in gene expression relative to sh empty. (d) Flow cytometry analysis of...

Additional file 3: Figure S3. LAMC2 expression correlates with CSC content and function. (a) qPCR analysis of LAMC2 gene expression in adherent cells versus spheres. Data are normalized to GAPDH and are presented as fold change in gene expression relative to adherent cells (indicated as Adh). (b) Western blot analysis of LAMC2 in adherent cells versus spheres. Parallel β-ACTIN immunoblotting was performed. (c) qPCR analysis for CD44, CD133 and L1CAM genes in adherent cells versus...

Additional file 4: Figure S4. Knockdown of LAMC2 does not affect cell growth. (a) Western blot analysis of LAMC2 in sh empty and LAMC2 knockdown cells. Parallel β-ACTIN immunoblotting was performed. (b) qPCR analysis for LAMC2 and CD44 gene expression in sh empty and LAMC2 knockdown cells. Data are normalized to GAPDH and are presented as fold change in gene expression relative to sh empty. (c) Representative images of sh empty and LAMC2 knockdown cells...

Additional file 6: Figure S6. Knockdown of LAMC2 affects tumorigenicity. (a) Representative images from a gelatin degradation assay of sh empty and LAMC2 knockdown cells. Nuclei were stained with Hoechst 33342 (blue), green represents actin (Alexa Fluor™ 488 Phalloidin) and red illustrates gelatin (Rodhamine). The white dashed line circles indicate the areas of degradation. (b) Degradation potential of sh empty and LAMC2 knockdown cells. (c) qPCR analysis for EMT genes in sh empty and LAMC2...

Additional file 7: Figure S7. Generation of LAMC2-EGFP knock-in human PDAC cells. (a) Flow cytometry for IRFP in PDAC cells 48 hours post-nucleofection. All cytometry gates were established based on isotype controls. (b) Flow cytometry for EGFP in PDAC cells 20 days post-nucleofection. All cytometry gates were established based on isotype controls. (c) PCR gDNA specific integration analysis. The positions of primers are indicated by arrows. (d) FACS profiles showing the expression of EGFP in...

Additional file 5: Figure S5. Loss of LAMC2 reduces stemness. (a) Flow cytometry for apoptotic cells as determined by AnnexinV/PI staining in control and LAMC2 knockdown cells. (b) Flow cytometry quantification of CD44 in sh empty and LAMC2 knockdown cells. (c) qPCR analysis of CD133 in sh empty and LAMC2 knockdown cells. Data are normalized to GAPDH and are presented as fold change in gene expression relative to sh empty. (d) Flow cytometry analysis of...