Fig. 4
From: Cell mediated ECM-degradation as an emerging tool for anti-fibrotic strategy
Methods to characterize ECM degradation ability of cells. A FITC-labeled substrate. The substrates are pre-labeled with fluorescent molecules (such as FITC) and then co-cultured with cells. Detection of areas without fluorescence can indirectly characterize the degradation ability of cells. B CHP staining. CHP can specifically bind denatured collagen and the degraded collagen can be stained with CHP conjugated to fluorescent molecules. C Gelatin zymography. Protein samples prepared from cells are separated by electrophoresis in a gel containing gelatin, and then MMPs in the samples degrade the gelatin at their respective sites, which are characterized by Coomassie Brilliant Blue staining. D HYP assay. The major proteins containing HYP are collagens. The HYP content of the supernatant, serum, or matrix co-cultured with cells can reflect the degree of degradation of the ECM. E qPCR. Real-time quantitative PCR (qPCR) can detect the expression of ECM-degrading related enzymes at the mRNA level, indicating the potential of cells to degrade the ECM. F Western blot. Using the specific combination of antigen and antibody, western blot can relatively qualitatively reflect the expression of ECM-degrading related enzymes at the protein level