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pmic7492-sup-0001-FigureS1.tif1651K

Figure S1. Biological and technical replicates showing calprotectin levels in lung increasing with bacterial burden.

Replicates of Figure 1, from left: PBS, 1.3×106 CFU, 8.4×107 CFU, 1.4×108 CFU, and 4.8×108 CFU. Ion intensity maps of S100A8, S100A9, and Histone H2A.1 are shown for each trial. (A) Shows a biological replicate of increasing bacterial burden, analyzed at 300 μm spatial resolution. (B) and (C) show technical replicates, analyzed at 500 μm spatial resolution.

pmic7492-sup-0002-FigureS2.tif1673K

Figure S2. Biological replicate showing calprotectin recruited to sites of infection.

Biological replicate of in-text Fig. 3, analyzed at 300 μm spatial resolution. (A) H&E stained serial sections of lung. (B) 40× magnification of areas with inflammation. (C) Ion intensity map of m/z 10,165, S100A8. (D) Ion intensity map of m/z 12,999, Sodiated S100A9. (E) Ion intensity map of m/z 14,050, Histone H2A.1.

pmic7492-sup-0003-FigureS3.tif1517K

Figure S3. Technical replicate showing calprotectin recruited to sites of infection.

Technical replicate of in-text Fig. 3, analyzed at 500 μm spatial resolution. (A) H&E stained serial sections of lung. (B) Ion intensity map of m/z 10,165, S100A8. (C) Ion intensity map of m/z 12,999, Sodiated S100A9. (D) Ion intensity map of m/z 14,050, Histone H2A.1.

pmic7492-sup-0004-FigureS4.tif1614K

Figure S4. Technical replicate showing calprotectin recruited to sites of infection.

Technical replicate of Supplemental Fig. 2, analyzed at 500 μm spatial resolution. (A) H&E stained serial sections of lung. (B) Ion intensity map of m/z 10,165, S100A8. (C) Ion intensity map of m/z 12,999, Sodiated S100A9. (D) Ion intensity map of m/z 14,050, Histone H2A.1.

pmic7492-sup-0005-FigureS5.tif1663K

Figure S5. PBS control lungs contain no appreciable signal for S100A8 or S100A9.

Lungs from mice mock-infected with 30 μl of PBS harvested at each time point, analyzed at 300 μm spatial resolution. (A) H&E stained serial sections of lung. (B) 40 × magnification of tissue. (C) Ion intensity map of m/z 10,165, S100A8. (D) Ion intensity map of m/z 12,999, Sodiated S100A9. (E) Ion intensity map of m/z 14,050, Histone H2A.1.

pmic7492-sup-0006-FigureS6.tif1170K

Figure S6. Average pixel intensity shows differences in protein abundance with time

Average Pixel intensity extracted from regions of interest from time course in Fig. 3 and control data in Supplementary Fig. 5. (A) m/z 10,165, S100A8. (B) m/z 12,999 Sodiated S100A9.

pmic7492-sup-0007-FigureS7.tif1723K

Figure S7. Ions of interest that scale with inflammation

Ion intensity maps extracted from the data show a number of species that seem to scale with infection. Images were analyzed at 300 μm spatial resolution of: (A) m/z 4,107, (B) m/z 10,165, S100A8, (C) m/z 10,225, (D) m/z 10,246, (E) m/z 10,384, (F) m/z 10,391, (G) m/z 13,207, (H) m/z 13.307.

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