Fig. 8
Prolonged IFN-γ signaling in Mmp12–/– primary peritoneal macrophages. Western blot analysis for pSTAT1-Y701 and STAT1 in primary peritoneal macrophages harvested from individual (a, e) Mmp12+/+ B10.RIII (n = 5 for each time point) and (b, d) Mmp12–/– B10.RIII (n = 4 for each time point) mice 4 days after induction of peritonitis. Cells were treated with 20 ng/mL of IFN-γ for 0–1080 min. c Ratios of pSTAT1-Y701 to STAT1 protein levels were determined after densitometry quantification of the western blots. The data are expressed as fold differences in the ratio of the means for Mmp12+/+ (n = 5 for each time point) and Mmp12–/– (n = 4 for each time point) B10.RIII mice. d Rescue of Mmp12–/– peritoneal macrophages with recombinant mouse MMP12 protein (1:100) incubated for the times shown (n = 4 for each time point). e Mmp12+/+ B10.RIII macrophages were incubated for 30 min with 100 nm specific MMP12 inhibitor Rxp470.1 before addition of 20 ng/mL IFN-γ (n = 4 for each time point). f Western blot analysis of IFN-γ, MMP12, iNOS, MHCII, S100A8, S100A9, STAT1, CD36, and STAT6 proteins in primary peritoneal macrophages harvested from Mmp12+/+ (n = 4) and Mmp12–/– (n = 4) B10.RIII mice at day 4 post-intraperitoneal injection with vehicle; and Mmp12+/+ B10.RIII mice treated daily with 5 mg/kg Rxp470.1 (n = 4) for 4 days during the induction of peritonitis. Actin or tubulin loading controls and molecular weight marker positions in all blots are as shown
