Journal: Cardiovascular Research

Article Title: STAT3-miR-17/20 signalling axis plays a critical role in attenuating myocardial infarction following rapamycin treatment in diabetic mice

doi: 10.1093/cvr/cvz315

Figure Lengend Snippet: Role of STAT3 in hearts of diabetic mice following rapamycin treatment. (A) Representative immunoblots for p-STAT3, total STAT3, and GAPDH expression in whole heart of C57 and db/db mice following 28 days of RAPA treatment. Densitometry analysis of immunoblots for the ratio of p-STAT3/STAT3 (*P < 0.05 vs. others) and the ratio of STAT3/GAPDH (*P < 0.05 vs. C57-Control; n = 3). (B) Myocardial infarct size following ischaemia/reperfusion in STAT3-deficient diabetic mice. High-fat diet (HFD)-fed WT and STAT3-deficient mice were treated with RAPA for 28 days prior to evaluation of I/R injury. Upper panel: representative images of heart sections following TTC staining (Scale indicates 5 mm). Lower panels: quantitative data of infarct size following I/R injury (*P < 0.001 vs. other) and rate-force product (*P < 0.01 vs. other; n = 5). (C) Isolated cardiomyocyte necrosis was determined by trypan blue staining following 40 min SI and 1 h RO (n = 4–8). (D) Representative pictures of TUNEL staining (scale indicates 100 µm) and quantitative data of cardiomyocyte apoptosis following 40 min SI and 18 h RO (n = 4; *P < 0.001 vs. controls, αP < 0.001 vs. both SI/RO and βP < 0.05 vs. WT-SI/RO). Statistics: one-way ANOVA.

Article Snippet: 23 Apoptosis was determined after 40 min SI and 18 h of RO using TUNEL staining kit (BD Biosciences, San Jose, CA, USA) as previously reported.

Techniques: Western Blot, Expressing, Staining, Isolation, TUNEL Assay

Journal: Experimental & Molecular Medicine

Article Title: A cell-penetrating peptide blocks Toll-like receptor-mediated downstream signaling and ameliorates autoimmune and inflammatory diseases in mice

doi: 10.1038/s12276-019-0244-0

Figure Lengend Snippet: a – d C57BL/6J male mice ( n = 8) were i.p. injected with PBS or 10 nmol/g of TIP1 1 h before i.p. administration of LPS (5 µg/g). Measurements of ( a ) TNF-α, ( b ) IL-12p40, and ( c ) IL-6 secretion were conducted on plasma samples after 2 h. d The protein levels of TNF-α and IL-6 were measured by western blotting in liver tissue. β-Actin was utilized as a loading control. Histograms represent the band intensities of TNF-α and IL-6 normalized to β-actin. e–l C57BL/6J male mice ( n = 5) were i.p. injected with PBS or 10 nmol/g TIP1 1 h before i.p. administration of LPS (5 µg/g). e , f Evaluation of TNF-α or IL-6 levels was performed by an ELISA on plasma samples after 24 h. g – j Levels of biological markers of renal dysfunction [blood urea nitrogen (BUN) and creatinine (Cr)] and markers of liver dysfunction [aspartate aminotransferase (AST) and alanine aminotransferase (ALT)] were measured in plasma samples. k Representative photographs of apoptotic cells in the kidney after TUNEL staining (scale bars represent 200 or 100 μm) and the quantitatively measured scores of TUNEL-positive cells in the histogram. l BALB/c male mice were i.p. injected with PBS or 10 nmol/g TIP1 1 h before i.p. administration of LPS (5 or 10 µg/g), and the survival rates were recorded for 5 days postinjection. The data shown represent at least three independent experiments ( n ≥ 3), and bars denote the mean ± SEM (* P < 0.05, ** P < 0.01). N.D.: not detected

Article Snippet: Apoptotic cells in the kidneys were quantified by staining them with a TUNEL Staining Kit (Merck Millipore, Billerica, MA, USA) and then examining them by confocal microscopy (LSM-700, Carl Zeiss Microscopy GmbH, Munich, Germany) using the Zen 2009 software package.

Techniques: Injection, Western Blot, Enzyme-linked Immunosorbent Assay, TUNEL Assay, Staining

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Gaseous signal molecule SO 2 regulates autophagy through PI3K/AKT pathway inhibits cardiomyocyte apoptosis and improves myocardial fibrosis in rats with type II diabetes

doi: 10.4196/kjpp.2022.26.6.541

Figure Lengend Snippet: (A) TUNEL staining of myocardial tissue of the rats in the Control, DC, DC + SO 2 and SO 2 groups (apoptosis-positive cells in TUNEL staining are in brown, and they are indicated by black arrows), n = 3. Images were acquired at 10 × 40 magnification; (B) Quantitative results of TUNEL-stained positive cells in the myocardial tissue of the rats in the respective group, **p < 0.01 vs. Control. ## p < 0.01 vs. DC; (C–G) Western blot detection of the expressions of Bax, Bcl2, Cleaved Caspase3, Caspase3, Cleaved Caspase9, Caspase9 in the myocardial tissue of the rats in the respective group. n = 3, *p < 0.05 vs. Control; # p < 0.05 vs. DC; (H, I) Detection of mRNA expression levels of Bax and Caspase3 in the myocardial tissue by RT-qPCR, **p < 0.01 vs. Control. ## p < 0.01 vs. DC, GAPDH is taken as the internal parameter. Values are presented as mean ± SD. SO 2 , sulfur dioxide; DC, diabetic cardiomyopathy.

Article Snippet: The fixed and embedded myocardial tissues were made into 4um thin sections, and the corresponding operations were performed in accordance with the instructions of the TUNEL staining kit (Keygene Biotech Co., Ltd.).

Techniques: TUNEL Assay, Staining, Control, Western Blot, Expressing, Quantitative RT-PCR

Journal: Theranostics

Article Title: Iron Oxide Nanoparticles Promote Cx43-Overexpression of Mesenchymal Stem Cells for Efficient Suicide Gene Therapy during Glioma Treatment

doi: 10.7150/thno.60160

Figure Lengend Snippet: In vivo therapeutic gains of using MFIONs-harbouring MSCs for suicide gene therapy against glioma. A) Schematic illustration to show the details of the therapeutic study. B) Survival curves of glioma rats after the different treatments. The differences between groups were compared with PBS group. C) Images of fixed brain samples and Nissl-stained brain sections. D) The areas of the gliomas were calculated with ImageJ according to the Nissl staining results. E) Pathological images of the cerebral sections (including H&E and TUNEL immunohistochemical staining). B) N.S.: no significant difference , **p < 0.01, compared to PBS group, based on Logrank. D) N.S.: no significant difference, **p < 0.01, based on one-way ANOVA. n = 5 for B). n = 3 for D). Data are means ± SD.

Article Snippet: A TUNEL immunohistochemical staining kit (ZK8004, ZSGB-BIO, Beijing, China) was performed to stain the brain samples following the manufacturer's protocol.

Techniques: In Vivo, Staining, TUNEL Assay, Immunohistochemical staining