Journal: International Journal of Molecular Sciences

Article Title: Epithelial PCSK6 Promotes Proliferation and Decreases Collagen Deposition by Fibroblasts Potentially via MMP Activation

doi: 10.3390/ijms27115104

Figure Lengend Snippet: PCSK6 promotes MMP activation. ( A ) Proteolytic activity of CM from mCherry-control, PCSK6-high and PCSK6-low A549 cells on the fluorogenic MMP substrate peptide {Mca}-Lys-Pro-Leu-Gly-Leu-{Dap(Dnp)}-Ala-Arg-NH2 ( n = 3). ( B ) Proteolytic activity of CM from mCherry-control, PCSK6-high and PCSK6-low A549 cells on the fluorogenic MMP14 substrate peptide MCA-PLA-C(OMeBz)-WAR(Dpa)-NH 2 ( n = 3). ( C ) Proteolytic activity of CM from mCherry-control, PCSK6-high and PCSK6-low A549 cells collected in the presence or absence of 50 µM furin convertase inhibitor (FC inh) on the fluorogenic MMP substrate peptide ( n = 3). ( D ) Proteolytic activity of CM from mCherry-control, PCSK6-high and PCSK6-low A549 cells collected in the presence or absence of 50 µM furin convertase inhibitor (FC inh) on the fluorogenic MMP14 substrate peptide ( n = 3). ( E – G ) Proteolytic activity of CM from mCherry-control, PCSK6-high and PCSK6-low A549 cells in the presence or absence of 500 µM broad-spectrum MMP inhibitor GM6001 ( E ), 50 µM Marimastat ( F ) or the combination of GM6001 and Marimastat ( G ). Fluorescence was measured at excitation 320/20 nm, emission 360/40 nm and normalized to the baseline fluorescence of the substrate peptide in the medium alone. MFI indicates mean channel fluorescence minus background. Data are expressed as mean ± SEM. Statistical significance was tested using mixed-effects analysis with Tukey’s post hoc test for multiple comparisons, and significance is shown relative to the indicated groups. p -values: * <0.05, ** <0.01, *** <0.001 and **** <0.0001.

Article Snippet: MMP activity was assessed using a fluorogenic MMP substrate peptide (CAS 720710-69-0, MCE, Monmouth Junction, NJ, USA), in the presence or absence of the broad-spectrum MMP inhibitor GM6001 (sc-203979, Santa Cruz Biotechnology) or Marimastat (CAS 154039-60-8, MCE).

Techniques: Activation Assay, Activity Assay, Control, Fluorescence