Studies on RNA integrity and gene expression in human myocardial tissue, pericardial fluid and blood, and its postmortem stability  Lucas González-Herrera, Aurora Valenzuela, Juan A. Marchal, José A. Lorente, Enrique Villanueva  Forensic Science International  Volume 232, Issue 1, Pages 218-228 (October 2013) DOI: 10.1016/j.forsciint.2013.08.001 Copyright © 2013 Elsevier Ireland Ltd Terms and Conditions

Fig. 1 mRNA extraction yield (ng mRNA/mg heart) (A), absorbance ratio 260/280 (A260/A280) (B), and RNA integrity number (RIN) (C) in five myocardial zones. Cardiac zones abbreviated as follows: ALV (anterior left ventricle); PLV (posterior left ventricle); IVS (interventricular septum); LRV (lateral right ventricle); and ARV (anterior right ventricle). The box represents interquartile range, and the vertical lines represent minimum and maximum values (adjacent values). Outliers are indicated by open circles, and are defined as values of over 1.5 and lower than 3 box lengths from the top end of the box. Extremes are indicated by cross, and are defined as values of over 3 box lengths from the top end of the box. *Significant correlation between RNA extraction yield and RIN in IVS (R2=0.4; P<0.05) (A) and (C). **Significant correlation among absorbance ratio in cardiac zones (R2=0.5–0.8; P<0.01) (B). ***Significant correlation among RIN values in cardiac zones (R2=0.4–0.7; P<0.01) (C). Forensic Science International 2013 232, 218-228DOI: (10.1016/j.forsciint.2013.08.001) Copyright © 2013 Elsevier Ireland Ltd Terms and Conditions

Fig. 2 TNNI3 (A), MYL3 (B), MMP9 (C), and TGFB1 (D) relative expression (2−ddCt) in heart tissue samples using different endogenous control genes: GAPDH, ACTB, TBP, and CYCA. Figures boxes and symbols are represented as explained in Fig. 1. Forensic Science International 2013 232, 218-228DOI: (10.1016/j.forsciint.2013.08.001) Copyright © 2013 Elsevier Ireland Ltd Terms and Conditions

Fig. 3 RIN values and GAPDH (A), TNNI3 (B), MYL3 (C) Ct values correlations in heart tissue. *Significant correlation between GAPDH Ct values and RIN (R2=0.5; P<0.05) (A). **Significant correlation between TNNI3 Ct values and RIN (R2=0.5; P<0.05) (B). ***Significant correlation between MYL3 Ct values and RIN (R2=0.5; P<0.05) (C). Forensic Science International 2013 232, 218-228DOI: (10.1016/j.forsciint.2013.08.001) Copyright © 2013 Elsevier Ireland Ltd Terms and Conditions

Fig. 4 TNNI3, MYL3, MMP9, TGFB1, and VEGFA relative expression (2−ddCt) in different cardiac zones (A), blood and pericardial fluid (B). aCardiac zones are abbreviated as listed in Fig. 1. Figures boxes and symbols are represented as explained in Fig. 1. Forensic Science International 2013 232, 218-228DOI: (10.1016/j.forsciint.2013.08.001) Copyright © 2013 Elsevier Ireland Ltd Terms and Conditions

Fig. 5 TNNI3 (A), MYL3 (B), MMP9 (C), TGFB1 (D), VEGFA (E) relative expression (2−ddCt) in myocardial tissue, blood and pericardial fluid in cases with ≤12h-PMI or >12h-PMI. Figures boxes and symbols are represented as explained in Fig. 1. *Significant differences in MYL3 gene expression in myocardial tissue and in **post-mortem blood (P<0.05) (B). ***Significant differences in MMP9 gene expression in pericardial fluid (P<0.05) (C). ****Significant differences in VEGF gene expression in blood (P<0.01) (E). Figures boxes and symbols are represented as explained in Fig. 1. *Significant differences in MYL3 gene expression in myocardial tissue and in **post-mortem blood (P<0.05) (B). ***Significant differences in MMP9 gene expression in pericardial fluid (P<0.05) (C). ****Significant differences in VEGF gene expression in blood (P<0.01) (E). Forensic Science International 2013 232, 218-228DOI: (10.1016/j.forsciint.2013.08.001) Copyright © 2013 Elsevier Ireland Ltd Terms and Conditions