Expression of connexin 43 protein in cardiomyocytes of heart failure mouse model

Figures

• 

  FIGURE 1. Myocardial characteristics of aortic valve disease. (A) HE staining of myocardial tissue sections. (B,C) Sirius red staining of myocardial tissue sections and its quantification analysis, fast green as background color, *P < 0.05; (D) immunofluorescence staining of Cx43 (red) and Troponin I (green) in the myocardium. Scale bar = 100 μm, AVS, Aortic Valve Stenosis; Con, Control; (E) remarkably fading of Cx43 and Troponin I fluorescent intensity were observed in AVS myocardium. *P < 0.05, **P < 0.01.

• 

  FIGURE 2. Generation of Cx43-BFP-GFP/ + reporter line. (A) Schematic diagram showing the targeting strategy of Cx43-BFP-GFP allele by homologous recombination using CRISPR/Cas9; (B) polymerase chain reaction (PCR) of genomic DNA, P1 + P2 means primer1 + primer2; P3 + P4 means primer3 + primer4; Heterozygous amplified a 564 bp band.

• 

  FIGURE 3. Generation and Characterization of Cx43-BFP-GFP/ + reporter line. (A,B) Whole-mount bright field and epifluorescence images of visceral organs from 6-weeks Cx43-BFP-GFP/ + mice; (C) immunostaining for BFP and Cx43 on heart, lung, brain, stomach, kidney, liver, spleen, and pancreas sections. The high magnification images of organs in areas labeled with the square dotted frame, respectively; (D) quantification of the labeling efficiency by the percentage of BFP+ Cx43+ in BFP+ cells. Data are the mean ± SD; n = 5 mice per group; (E) quantification of the labeling specificity by the percentage of BFP+ Cx43+ in Cx43+ cells. Data are the mean ± SD; n = 5 images per group. Yellow scale bar = 1 mm, white scale bar = 100 μm.

• 

  FIGURE 4. Characterization of Cx43-BFP-GFP/ + reporter line of E16.5 mice. (A) Whole-mount bright field and epifluorescence images of E16.5 Cx43-BFP-GFP/ + mice; (B) whole-mount bright field and epifluorescence images of visceral organs from E16.5 Cx43-BFP-GFP/ + mice; (C) immunostaining for BFP and Cx43 on the sections of several organs. The high magnification images of organs in areas labeled with the square dotted frame, respectively; (D) quantification of the labeling efficiency by the percentage of BFP+ Cx43+ in BFP+ cells. Data are the mean ± SD; n = 5 images per group; (E) quantification of the labeling specificity by the percentage of BFP+ Cx43+ in Cx43+ cells. Data are the mean ± SD; n = 5 images per group. Yellow scale bar = 1 mm, white scale bar = 100 μm.

• 

  FIGURE 5. Expression of Cx43 in different cells on 6-week Cx43-BFP-GFP/ + mice. (A) Immunostaining for BFP, Cdh5, CD45, α-SMA, and Troponin I on heart sections; (B–D) immunostaining for BFP, α-SMA, PDGFRα, Cdh5, and CD45 on liver, kidney and spleen sections. The high magnification images of organs in areas labeled with the square dotted frame. Scale bar = 100 μm.

• 

  FIGURE 6. Expression of Cx43 in cardiomyocytes of heart failure. (A) Representative M-mode echocardiograms in the control and TAC groups; EF (B) and FS (C), LVIDd (D), and LVIDs (E) changes in TAC mice evaluated by echocardiography, *P < 0.05, ***P < 0.001, ****P < 0.0001; (F) whole-mount bright field images of the heart in two groups; (G) HE staining of heart sections of normal mice and TAC-operated mice. (H,I) Sirius red staining of the heart sections of normal mice and TAC-operated mice and its quantification analysis, fast green as background color, *P < 0.05; (J) expression of BFP (red) in the myocardium of Cx43-BFP-GFP mice with heart failure. Red scale bar = 1 mm, black scale bar = 200 μm, white scale bar = 100 μm; (K) fading of BFP fluorescent intensity was observed in TAC mice, *P < 0.05.