Journal: Cardiovascular diabetology

Article Title: Tenascin-C drives cardiovascular dysfunction in a mouse model of diabetic cardiomyopathy.

doi: 10.1186/s12933-025-02780-y

Figure Lengend Snippet: Fig. 5 Apoptosis and CD68+ expression. A Representative images of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) staining for apoptotic cells in the left ventricular wall of wild-type (WT) and Tenascin-C knockout (TNC-KO) mice, diabetic vs. non-diabetic. TUNEL stain ing was performed to assess the extent and distribution of apoptotic cells in the cardiac tissue. Green dots represent apoptotic cells resulting from the mixture of TUNEL-stained and DAPI-counterstained nuclei. Blue dots represent non-apoptotic cells stained with DAPI only. No evidence of apoptosis was found in WT non-diabetic and TNC-KO non-diabetic heart tissue, whereas apoptotic cells were frequently found in areas thought to represent remodel ing, visible as green staining in WT diabetic mice. Fewer TUNEL-positive cells were found in TNC-KO diabetic heart tissue compared to diabetic WT ani mals. Scale bar = 100 µm (200× magnification). B Immunoperoxidase staining for CD68. Representative images of myocardial tissue from WT and TNC-KO mice, diabetic vs. non-diabetic, showing CD68+ macrophages in the myocardium (magnification× 200, scale bar: 50 μm). (C) No statistical difference in numbers of CD68+ macrophages in the myocardium between the groups. The graph shows the number of CD68-positive cells. Data are expressed as mean ± SD; n = 4/group. (D)High glucose and (E)Tenascin C-induced apoptosis in H9C2 cells. Representative fluorescence pictures demonstrate the pres ence of positive TUNEL and DAPI as well as merged staining in positive control (cells were treated with 500 U/mL DNase I), negative control (cells were with the enzymatic solution without the labeled antibody), control, glucose (30 mM) and rh TNC (5 µg/mL) treatment groups. (G-H) Quantitative analysis of the TUNEL assay. DAPI, 4′,6-diamidino-2-phenylindole; Rh TNC, recombinant human Tenascin-C; TNC-KO, Tenascin-C knockout; TUNEL, terminal deoxy nucleotidyl transferase-mediated dUTP nick-end labelling; WT, wildt-type. Data shown as mean ± SD, n = 3–6/ condition; *p < 0.05

Article Snippet: Afterwards, the coverslips were washed 3 × with PBS for 5 min. Fluorescence labelled secondary antibody (Texas Red TI-1000, Vector, 1:200), diluted in PBS, was added and incubated for 1 h at RT under light protection, in the dark.

Techniques: Expressing, TUNEL Assay, Staining, Knock-Out, Immunoperoxidase Staining, Fluorescence, Positive Control, Negative Control, Labeling, Control, Recombinant