Supplementary Data

• Supplementary Figure 1 - qPCR (A) and western blot (B) analyses of cardiac tissue revealed a complete absence of 11βHSD1 enzyme expression in HSD1 null mice. *** P<0.005 n=4/4 (C) Levels of troponin were measured in the plasma of mice 1 day post-MI. n=5/5 (PDF 138 KB)
• Supplementary Figure 2 - Gating strategy for flow cytometry analysis of blood neutrophils. Blood cells are gated by granularity and size (by SSC- Side Scatter and FSC, forward scatter) with counting beads added to ascertain absolute cell numbers (A). Singlets are selected to exclude cell clumps (B; FSC- H/FSC-A; - height/-area). CD45+ cells are selected (C) and of these, neutrophils are the CD11b + Ly6G+ cells (D). (PDF 149 KB)
• Supplementary Figure 3 - Chemokine receptor protein expression in neutrophils was measured by flow cytometry (gating strategy defined in Supplementary Figure 2). CXCR4 expression, as calculated by mean fluorescence intensity (MFI) value, was lower in bone marrow of Hsd11b1-/- neutrophils 1 day post-MI than WT (A; P<0.05). CXCR2 expression was similar on bone marrow neutrophils between the two groups (C). Expression of the retention factor Cxcl12 in the bone marrow of WT and Hsd11b1-/- animals post-MI was measured by qPCR (B; n=4/3). (PDF 135 KB)
• Supplementary Figure 4 - Characterisation of cell types isolated from fibroblast enrichment. qPCR analysis of RNA from isolated fractions 1-3. (PDF 50 KB)
• Supplementary Figure 5 - To determine the response in cardiac fibroblasts under “normal” conditions qPCR analysis of Cxcl2 (A) and Cxcl5 (B) was performed on RNA isolated from the cellular fractions 2 and 3 (Ft2/3) in wt mice. Values were relative to whole infarct tissue and normalised to GAPDH **p<0.01, ***P<0.005, n=5-6 per group. (PDF 107 KB)
• Supplementary Figure 6 - Hsd11b1 gene expression is detected by PCR in Thy1^+ve and Thy1^-ve fractions of the DDR^+ve, Col1a1^+ve mouse cardiac fibroblast population, isolated from 3 separate mouse hearts, 24h after induction of myocardial infarction by coronary artery ligation. The bands represent a 587bp product corresponding to murine Hsd11b1. Primers were Forward: 5’-GTCCCTGTTTGATGGCAG-3, and Reverse: 5’-AGGATCCAAGCAAACTTGCTTGCA-3. (PDF 322 KB)