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Blood, Vol. 107, Issue 12, 4946-4953, June 15, 2006
Rapid ATP-induced release of matrix metalloproteinase 9 is mediated by the P2X7 receptor
Blood Gu and Wiley
107: 4946
Supplemental materials for: Gu and Wiley
Files in this Data Supplement:
- Table S1. MMP-9 spontaneous release in subsets of mononuclear cells (PDF, 34.1 KB) -
Four subsets of mononuclear cells from healthy subjects were separated using MACS magnetic beads. Purified cells from each subset were counted and resuspend in NaCl medium at 2×107/mL with 1mM Ca2+ at 37°C. After 60 min supernatants were collected and MMP-9, TIMP-1 concentrations were determined as “Basal” (ng/mL) using ELISA. Proportion of MMP-9 contribution from each single subset (protein concentration multiplies by volume of cell suspension) versus total amount of released MMP-9 from all four subsets was calculated and presented as “%.”
- Figure S1. Rapid release of MMP-9 and TIMP-1 by PMA (JPG, 82.2 KB)
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Human mononuclear cells (2x107/mL) were incubated with PMA for 15 min at 37°C in NaCl medium containing 1 mM CaCl2. Supernatant and cell pellet were collected by centrifugation. Cell pellets were lysed in original volume. MMP-9 activity in supernatants (a) and cell pellet lysates (b) was detected by gelatin zymography. MMP-9 (c) and TIMP-1 (d) concentration in both supernatants and cell pellet lysates were determined by ELISA. One representative experiment of two.
- Figure S2. Inhibition of ATP or BzATP-induced rapid MMP-9 release by OxATP (JPG, 57 KB)
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Mononuclear cells (2×107/mL) were incubated with OxATP (300 µM) at 37°C in NaCl medium containing 1 mM CaCl2 for 60 min, followed by addition of various concentration of ATP or BzATP as indicated. Supernatants were collected after 30 min. MMP-9 and TIMP-1 concentrations (ng/mL) were measured by ELISA. Curves are representative data from two healthy subjects with wild type P2X7 function who have been studied in Figure 3.
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