Extended Data Fig. 7: TRAIL is upregulated by cIAP2−/− mice post-MI.
From: Hematopoietic expression of cIAP2 drives inflammation and heart failure after myocardial infarction
a) mRNA expression of Tnfa, Il1b, and IL6 normalized to HPRT from Day 7 post-MI WT or cIAP2−/− spleens. N = 3 per treatment group. b) Immunoblotting of splenocyte lysates from 7-day post-MI from WT or cIAP2−/− mice were immunoblotted for TRAIL (top) and TNFα, cleaved (active) Caspase 8 (p41 and p18), cleaved Caspase 3 (p17/19), Annexin A1 or vinculin. Imaging representative of two similar experiments, respectively. c) (Top panel) TRAIL and IRF1 protein expression from bone marrow cells collected from WT and cIAP2−/− mice 24-hours post-MI. (Bottom panel) Expression of TRAIL compared between dendritic cells (CD11c+) isolated from WT, cIAP2−/− and cIAP1−/− bone marrow 24-hours post-MI. d) High-resolution immunofluorescence microscopy of spleen sections detecting TRAIL and DR5 in cIAP2−/− mice 3 days post-MI; arrowheads indicate increased colocalized staining of TRAIL on cell surface of BST2+ splenocytes. Scale bar = 25 μm; images representative of three animals per treatment. e) Increased LVESD 28 days post-MI in both WT and cIAP2−/− MI recipient mice after anti-TRAIL (N2B2) antibody treatments. Control IgG2 recipients: N = 3 sham operated controls (WT); N = 4 WT and cIAP2−/− mice per MI treatment group. Anti-TRAIL recipients: N = 3 sham operated controls (WT); N = 5 WT and cIAP2−/− mice per MI treatment group. f) Flow cytometry of inflammatory cells from hearts (left panels) and spleens (right panels) of MI recipient mice following TRAIL neutralizing antibody treatment. cIAP2−/− mice exhibit more pronounced increases in cardiac tissues. Control IgG2 recipients: N = 3 sham operated controls (WT); N = 4 WT and cIAP2−/− mice per MI treatment group. Anti-TRAIL recipients: N = 3 sham operated controls (WT); N = 5 WT and cIAP2−/− mice per MI treatment group. g) Schematic summary of proposed activity of splenic cIAP2−/− pDCs following TNFα or death-promoter TNFSF ligand (TRAIL) signaling (via TNFR1 or DR5, respectively). Error bars denote Mean +/- SD; p values were calculated using two-sided t test (a), two-way ANOVA with Tukey’s test (e) or two-way ANOVA with Bonferroni’s correction (f).
