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Fig. 3 | Journal of Biomedical Science

Fig. 3

From: TRIM18 is a critical regulator of viral myocarditis and organ inflammation

Fig. 3

Deletion of TRIM18 protects mice from myocarditis induced by RNA virus CVB3 in vivo. a Hematoxylin and eosin (H&E)-staining of heart sections from age-matched Trim18+/+ (WT) and Trim18−/− (KO) male mice after intraperitoneal infection with CVB3 (1 × 107 PFU per mouse) for 4 days. Scale bars represent 1000 μm for original images and 200 µm for enlarged images. b Histology score analysis of viral myocarditis in heart sections from mice as in (a). c Immunoblot (IB) analysis of TRIM18 expression in hearts from WT mice without or with intraperitoneal CVB3 acute infection (1 × 107 PFU per mouse) or chronic infection (1 × 103 PFU per mouse) for 2 days. The position of protein markers (shown in kDa) is indicated at right. d Immunohistochemistry (IHC) analysis of TRIM18 and TRIM21 expression in hearts from WT and KO male mice after CVB3 infection. e IHC analysis of the infiltrated cells in CVB3 infected hearts using anti-macrophage marker antibody, anti-neutrophil marker antibody, anti-NK cell marker antibody and anti-T cell marker antibody, respectively. Scale bars represent 100 μm in (d, e). f Representative M-mode images of hearts from WT and KO male mice at day 4 after CVB3 infection by echocardiography analysis. g–h Cardiac function analysis of ejection fraction (EF) g and fractional shortening (FS) h of hearts from mice as in (f) (n = 5 per group). i The assessment of heart weight/baseline body weight from WT and KO male mice (n = 5 per group) at day 0 or day 6 after CVB3 infection. j The qRT-PCR analysis of brain natriuretic peptide (BNP) mRNA in the heart of from WT and KO male mice (n = 5 per group) at day 1, day 2 or day 4 after CVB3 infection.; results are presented relative to those of mock mice. k Survival of age-matched WT and KO male mice after intraperitoneal infection with CVB3 (1 × 107 PFU per mouse) (n = 10 per group). l Viral titers in homogenates of hearts from WT and KO male mice at day 2 (D2), day 5 (D5) and day 14 (D14) after CVB3 infection (n = 5 per group for D2 and D5, n = 3 per group for D14). m–q, ELISA of IFN-α (m), IFN-β (n) IL-6 (o), TNF-α (p) and IL-1β q in hearts from mice as in k (n = 5 per group). Error bars indicate standard error of the mean for results in (b, g–j, l–q). NS, not significant (p > 0.05), **p < 0.01, ***p < 0.001, and p value was calculated by unpaired two-tailed Student’s t test and Gehan-Breslow-Wilcoxon test for survival analysis. Data are representative of three independent experiments

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