Supplementary table I. Real-time primers used in the study. The fold change was obtained by normalizing the gene expression number to those of HPRT, then comparing the samples to untreated or naive mice.
Supplementary table II. Colocalization analysis for images in figure 6, Pearson s correlation coefficient and % of MMP13 colocalized with F4/80, SMA and Gr1.
Supplementary figure S1. C57BL/6 (WT) and mmp12 -/- mice were infected with approximately 30 infective cercariae of S. mansoni and then euthanized on wk 9. Liver tissues were fixed in Bouin-Hollande solution and stained with Giemsa. Number of Eosinophils (a) and mast cells (b) were counted in liver granulomas. Hydroxyproline levels normalized to worm pairs (c) are shown. Statistical significance is measured using unpaired student s t test (n=14/group). The data are combined from two independent experiments that produced similar results.
Supplementary figure S2. C57BL/6 (WT) and mmp12 -/- mice were infected with 30 infective cercariae of S. mansoni and euthanized on wk 12. Liver tissues were fixed in Bouin-Hollande solution and stained with Giemsa. Average granuloma volumes and liver hydroxyproline levels normalized to egg count are shown. Statistical significance is measured using unpaired student s t test (n=14/group). The data are combined results from two independent experiments that produced similar results. Supplementary figure S3. Col 1a, Col 3a and Col 6a expression in liver tissues at 9 wk infection with S. mansoni. Values were normalized to hypoxanthine phosphoribosyltransferase (hprt), and fold changes in WT and mmp12 -/- were generated by comparing with WT and mmp12 -/- unchallenged mice, respectively. The data show the means ± SEM. Statistical significance of data was determined using one-way ANNOVA (n=5/group)
Supplementary figure S4. C57BL/6 (WT) and mmp12 -/- mice were infected with approximately 30 infective cercariae of S. mansoni and euthanized after 9 weeks. Liver tissues were fixed in Bouin-Hollande solution and stained with Giemsa. Numbers of eosinophils were counted in liver granulomas. The data show the means ± SEM.
Supplementary figure S5. WT and mmp12 -/- mice were sensitized and challenged with S. mansoni eggs. On day 8 post-challenge, bronchoalveolar lavage (BAL) fluid was recovered. Numbers of cells in the BAL fluid were plotted as means ± SEM. BAL fluids were analyzed for IL-13 using a multipex cytokine analysis method. The data show the means ± SEM. One of two independent experiments are shown. Supplementary figure S6. WT and mmp12 -/- mice were sensitized and challenged with S. mansoni eggs. On day 8 post-challenge, real-time PCR analysis was performed on 50 mg lung samples obtained from WT and mmp12 -/- mice. Genes analyzed: tgf-β and egr-1 as described in methods.
Supplementary figure S7. WT and mmp12 -/- mice were sensitized and challenged with S. mansoni eggs. On day 8 post-challenge, real-time PCR analysis was performed on 50 mg lung samples obtained from WT and mmp12 -/- mice. Genes analyzed include col 1a, Col 3a and col 6a as described in methods. Supplementary figure S8. BMDM were seeded at a concentration of 0.5 10 6 per well in 24- well plates and stimulated with IL-4, IL-13, IL-21, IFN-γ and TNF-α at a concentration of 20 ng/ml for 16 h. Real-time PCR analysis was performed for mmp12 and mmp13 and values were normalized to hprt and compared to untreated cells. The data show the means ± SEM.
Supplementary figure S9. Thioglycollate-elicited mouse peritoneal macrophages were seeded at a concentration of 0.5 10 6 per well in 24-well plates and stimulated with IL-4 (20 ng/ml) in the presence and absence of anti-il4rα, anti-il2γc, or isotype control Ab for a period of 16 h. Real-time PCR analysis was performed for mmp13 expression and values were normalized to hprt and compared to untreated cells. The data show the means ± SEM. Similar results were obtained in two independent experiments.
Supplementary figure S10. Thioglycollate-elicited mouse peritoneal macrophages were seeded at a concentration of 0.5 10 6 per well in 24-well plates and stimulated with IL-13 (20 ng/ml) in the presence and absence of anti-il4rα, anti-il2γc, or isotype control Ab for a period of 16 h. Real-time PCR analysis was performed for mmp13 expression and values were normalized to hprt and compared to untreated cells. The data show the means ± SEM. Similar results were obtained in two independent experiments.
Supplementary figure S11. C57BL/6 (WT) and mmp12 -/- mice were infected with 30 infective cercariae of S. mansoni and euthanized after 9 weeks. Liver tissues were fixed in Bouin-Hollande solution and stained with Giemsa. Macrophages present at the center or periphery of the granuloma as well as the total number were enumerated. The data show the mean ± SEM. The data are combined results from two independent experiments that produced similar results.
Supplementary figure S12. S. mansoni egg challenged lungs from wild type and mmp12 -/- mice were fixed in Bouin-Hollande solution and stained with Giemsa. Macrophages present at the center or periphery of the granuloma as well as the total number were enumerated. The data show the means ± SEM. The data are combined results from two independent experiments that produced similar results.