Fig 1: Functional and structural changes to eCTs 14 days after exposure to neutron radiation. A) Schematic of the platform with eCT tissue being characterized. B) Histological staining of eCTs using a canonical marker for cardiomyocytes (a‐actinin) and fibroblasts (vimentin). C) Cardiac troponin T (cTnT) concentration in the supernatant of eCT compartments over the duration of the experiment, normalized to control groups, with significance shown by two‐way ANOVA with multiple comparisons. D) Representative bright field image of eCTs at endpoint. E–G) Characterization of contractile function E), tissue morphology at rest F) at the 14‐day endpoint. Each tissue is normalized to itself at baseline and then normalized to the average value of the control group. G) Characterization of excitability of cardiac muscle tissues, normalized to the average value of the control group. Statistical significance was determined by one‐way ANOVA with Welch's correction and corrections for multiple comparisons (C,E,F) or by the Student's t‐test (G) (p‐values not shown on plots are p > 0.1).
Fig 2: Overexpression of TLR4 abolished the inhibitory effect of silenced ALKBH5 on myocardium damage by radiation in mice induced. (a) Transfection efficiency was measured by qPCR. (b)–(d) The cardiac function including LVEF, LVPW-d, and LVPW-s was analyzed using the ultrasound imaging system. (e) and (f) The levels of cTnT and BNP (B-type natriuretic peptide) in the serum were detected using an ELISA kit. (g) Myocardial damage was visualized using the HE staining. All data are expressed as the mean values ± SD. n = 6 independent biological replicates/group in animal experiments.
Fig 3: Downregulation of ALKBH5 reversed the myocardium damage induced by radiation in mice. (a) Transfection efficiency was measured by qPCR. (b)–(d) The cardiac function including LVEF, LVPW-d, and LVPW-s was analyzed using the ultrasound imaging system. (e) and (f) The levels of cTnT and BNP (B-type natriuretic peptide) in the serum were detected using an ELISA kit. (g) Myocardial damage was visualized using the HE staining. All data are expressed as the mean values ± SD. n = 6 independent biological replicates/group in animal experiments.
Fig 4: RIHD and upregulated ALKBH5 expression in RIHD mice. (a)–(c) The cardiac function including LVEF, LVPW-d, and LVPW-s was analyzed using the ultrasound imaging system. (d) and (e) The levels of cTnT and BNP (B-type natriuretic peptide) in the serum were detected using an ELISA kit. (f) Myocardial damage was visualized using the HE staining. (g)–(i) The levels of ALKBH5 were examined using qPCR, IHC assays, and western blot in the heart tissues of mice, respectively. All data are expressed as the mean values ± SD. NS: no significance. n = 6 independent biological replicates/group in animal experiments.
Fig 5: Tissue pre-conditioning impacts extent of cellular damage from reperfusion injury. Cellular damage 24 h following reperfusion is quantified by LDH (A,B) and cardiac troponin (cTnT) (C,D) release in tissues subjected to ischemia compared to their normoxic controls for all conditioning groups, or normalized to their respective controls (B,D). Cellular damage 72 h following reperfusion is similarly quantified by LDH release in (E,F), and cTnT release in (G,H), respectively. Data shown is from n = 3-6 tissue replicates and mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 and statistical analysis was performed using ordinary one-way ANOVA corrected for multiple comparisons using Tukey’s hypothesis testing.
Supplier Page from Abcam for Human Cardiac Troponin T ELISA Kit