Journal: Bio-protocol
Article Title: Non-Enzymatic Isolation of Cancer-Associated Fibroblasts From Human Prostate Tumor Explants
doi: 10.21769/BioProtoc.5614
Figure Lengend Snippet: . The results indicate that collagen type I (col1a1) and vimentin are expressed in both healthy and cancer-associated fibroblasts (HPFs and CAFs), confirming their mesenchymal identity. Conversely, α-smooth muscle actin (α-SMA) is detected exclusively in CAFs, reflecting their activation compared to the HPFs. As expected, CD31 is exclusively expressed in endothelial cells (ECFCs), while E-cadherin and claudin are only detected in epithelial cells (DU145), in accordance with their role in epithelial junctions. HSP90 was used as a loading control to ensure equal protein content across samples. HPF and CAF were derived from three different explants. Note that α-SMA expression differs among the three CAF samples, reflecting inter-patient biological variability.
Article Snippet: Human epithelial PCa cell line DU145 (RRID: CVCL 0105) obtained from ATCC and routinely tested for Mycoplasma contamination using the MycoAlert Mycoplasma Detection kit (Lonza, #LOLT07710) 4.
Techniques: Activation Assay, Control, Derivative Assay, Expressing