Journal: Respiratory Research

Article Title: An improved precision-cut lung slices technique for pulmonary vasoconstriction

doi: 10.1186/s12931-026-03600-x

Figure Lengend Snippet: Acute hypoxia experiment in PCLS. A , Blood vessel morphology in the control group was recorded under the bright-field microscope at the beginning of perfusion, and 40 mM potassium chloride solution was added after the normoxic perfusion was completed. And the subsequent immunofluorescence staining was conducted to determine CX40 expression (right). B , The vascular morphology image in the hypoxic group was recorded in the same way while hypoxic perfusion solution with 1% oxygen was used at the fifth minute after the normmoxic perfusion, and the final exposure to 40 mM potassium chloride solution. And the subsequent immunofluorescence staining was conducted to determine CX40 expression (right). The vessel area was marked with a red dashed line; scale bar: 100 μm (A and B). Due to coverslip mounting required for immunofluorescence staining, the vascular morphology in 200 μm-thick PCLS exhibited slight morphological changes (A and B, right). C , The statistical comparisons of the alteration of vascular area recorded every minute during the entire 35-minute perfusion process, n=4 for each

Article Snippet: Gelatin (porcine skin, type A, 300 g Bloom, Vetec), Cell Tracker CM-DiI (Yeasen Biotechnology, 40718ES50), low melting point agarose (CAS 9012-36-6), tissue perforator (GLOBALEBIO), Penicillin-Streptomycin (1%, Gibco), WST-1 Cell Proliferation and Cytotoxicity Assay Kit (Biyuntian, C0035), CX40 Polyclonal Antibody (BA1593, Boster, rabbit polyclonal antibody), FITC-conjugated Goat Anti-Rabbit IgG H&L (Servicebio, GB22303), α-SMA specific antibody (Proteintech, 67735-1-Ig, Clone: 1E9A11), Cy3-conjugated Goat anti-Mouse IgG H&L (Proteintech, SA00009-1), and Antifade Mounting Medium (Servicebio, G1401).

Techniques: Control, Microscopy, Immunofluorescence, Staining, Expressing

Journal: Respiratory Research

Article Title: An improved precision-cut lung slices technique for pulmonary vasoconstriction

doi: 10.1186/s12931-026-03600-x

Figure Lengend Snippet: Immunofluorescence and H&E staining images of pulmonary artery and vein in consecutive paraffin sections. A , CX40 (green) and α-SMA (red) were expressed in artery, whereas only α-SMA (red) in vein. B , Venous and arterial tissue demonstrated in hematoxylin-eosin staining in mouse lung tissue sections. Scale bar: 50 μm

Article Snippet: Gelatin (porcine skin, type A, 300 g Bloom, Vetec), Cell Tracker CM-DiI (Yeasen Biotechnology, 40718ES50), low melting point agarose (CAS 9012-36-6), tissue perforator (GLOBALEBIO), Penicillin-Streptomycin (1%, Gibco), WST-1 Cell Proliferation and Cytotoxicity Assay Kit (Biyuntian, C0035), CX40 Polyclonal Antibody (BA1593, Boster, rabbit polyclonal antibody), FITC-conjugated Goat Anti-Rabbit IgG H&L (Servicebio, GB22303), α-SMA specific antibody (Proteintech, 67735-1-Ig, Clone: 1E9A11), Cy3-conjugated Goat anti-Mouse IgG H&L (Proteintech, SA00009-1), and Antifade Mounting Medium (Servicebio, G1401).

Techniques: Immunofluorescence, Staining

Journal: Respiratory Research

Article Title: An improved precision-cut lung slices technique for pulmonary vasoconstriction

doi: 10.1186/s12931-026-03600-x

Figure Lengend Snippet: Immunofluorescence and H&E staining images of consecutive paraffin sections. A , CX40 (green) and α-SMA (red) were expressed in both pulmonary artery and bronchus. B , bronchus and arterial tissue structures demonstrated by hematoxylin-eosin staining in mouse lung tissue sections. Scale bar: 50 μm

Article Snippet: Gelatin (porcine skin, type A, 300 g Bloom, Vetec), Cell Tracker CM-DiI (Yeasen Biotechnology, 40718ES50), low melting point agarose (CAS 9012-36-6), tissue perforator (GLOBALEBIO), Penicillin-Streptomycin (1%, Gibco), WST-1 Cell Proliferation and Cytotoxicity Assay Kit (Biyuntian, C0035), CX40 Polyclonal Antibody (BA1593, Boster, rabbit polyclonal antibody), FITC-conjugated Goat Anti-Rabbit IgG H&L (Servicebio, GB22303), α-SMA specific antibody (Proteintech, 67735-1-Ig, Clone: 1E9A11), Cy3-conjugated Goat anti-Mouse IgG H&L (Proteintech, SA00009-1), and Antifade Mounting Medium (Servicebio, G1401).

Techniques: Immunofluorescence, Staining

Journal: bioRxiv

Article Title: Cell-autonomous Wnt activity promotes transient re-programming and cell cycle re-entry of coronary artery endothelial cells

doi: 10.64898/2026.02.23.707374

Figure Lengend Snippet: ( A ) Experimental design to assess expression of VegfR2 and endomucin in single artery endothelial cells (saECs), post-MI. ( B , C ) Quantification of ( B ) VegfR2 and ( C ) Endomucin, in saECs, post-MI. p<0.0001. Each data point is a TdTomato + single artery cell. N=3 and N=5 hearts were quantified for VegfR2 and Endomucin expression, respectively. ( D ) Confocal images of saECs in Cx40CreER; Rosa26 TdTomato ; Cx40 eGFP/+ hearts, post-MI, showing TdTomato + saECs in red and expression of eGFP as a reporter for Cx40 expression in green. Arrowheads point to TdTomato + saECs with low levels of eGFP signal, as compared to the artery tip. ( E , H ) Confocal images of post-MI watersheds, showing TdTomato + saECs in red and immunostaining for VegfR2 in green in ( E ) control ( Cx40CreER; Rosa26 TdTomato ) and ( F ) knockout ( Fzd4 L/L ; Cx40CreER; Rosa26 TdTomato ) MI hearts. F , G are insets from E and show TdTomato + saECs express VegfR2 (orange arrowheads). I , J are insets from H and show reduced expression of VegfR2 by TdTomato + saECs (magenta arrowheads). ( K , N ) Confocal images of post-MI watersheds, showing TdTomato + saECs in red and immunostaining for Endomucin in green in ( K ) control ( Cx40CreER; Rosa26 TdTomato ) and ( N ) knockout ( Fzd4 L/L ; Cx40CreER; Rosa26 TdTomato ) MI hearts. L , M are insets from K and show TdTomato + saECs express Endomucin (orange arrowheads). O , P are insets from N and show reduced expression of Endomucin by TdTomato + saECs (magenta arrowheads). Scalebars: 50µm. saECs, single artery endothelial cells; P, postnatal day; KO, knockout; cap, capillary; MI, myocardial infarction.

Article Snippet: Following mouse lines, Cx40CreER , ApjCreER , Cdk1 fl/fl (The Jackson Laboratory, 129S(B6N)-Cdk1 tm1Eddy /J, stock number 028028), Fzd4 fl/fl (The Jackson Laboratory, B6;129-Fzd4 tm2.1Nat /J, stock number 011078), Wls fl/fl (The Jackson Laboratory, Wls tm1.1Whsu /J, stock number 027484), Rosa26 TdTomato Cre reporter line (The Jackson Laboratory, B6.Cg-Gt[ROSA]26Sortm9[CAG-TdTomato]Hze/J, stock number 007909), Cx40 eGFP/+ and Axin2-d2EGFP were used in the experiments.

Techniques: Expressing, Immunostaining, Control, Knock-Out

Journal: Biomolecules

Article Title: Trandolapril Attenuates Pro-Arrhythmic Downregulation of Cx43 and Cx40 in Atria of Volume Overloaded Hypertensive and Normotensive Rats

doi: 10.3390/biom15101457

Figure Lengend Snippet: Protein levels of Cx43 ( A ) and Cx40 ( B ) normalized to GAPDH assessed by Western blot analysis. Hypertensive TGR rats exhibited lower levels of Cx43 and Cx40 in both the left and right atria compared to normotensive rats. A pronounced decrease in Cx43 and Cx40 protein levels was observed in both atria of normotensive HSD and hypertensive TGR rats due to volume overload. Treatment with an ACEi increased Cx43 and Cx40 levels ( p > 0.05) in both atria of volume-overloaded normotensive and hypertensive rats. In contrast, ARB treatment increased Cx43 but did not affect Cx40 levels in either atrium of volume-overloaded rats. HSD—Hannover Sprague Dawley rats; TGR—Ren-2 transgenic rats; ACF—aortocaval fistula, surgical model of volume overload; ACEi—treatment with the angiotensin-converting enzyme inhibitor, trandolapril; ARB—treatment with an angiotensin II type 1 (AT 1 ) receptor blocker, losartan. n = 10 per group. Data are presented as means ± SD; a p < 0.05 vs. HSD, b p < 0.05 vs. HSD ACF, c p < 0.05 vs. TGR, d p < 0.05 vs. TGR ACF. Western blot original images can be found in .

Article Snippet: Membranes were blocked in 5% low-fat milk ( w / v in TBS-T) for 4 h at room temperature and incubated overnight at 4 °C with the appropriate primary antibodies: anti-total Cx43—1:5000, C6219, Sigma-Aldrich, MI, USA; anti-Cx40—1:2000, sc-365107, Santa Cruz Biotechnology, Dallas, TX, USA; anti-PKCε—1:1000, sc-214, Santa Cruz Biotechnology, TX, USA; anti-PKCδ—1:1000, sc-213, Santa Cruz Biotechnology, Dallas, TX, USA; anti-MMP-2—1:1000, sc-10736, Santa Cruz Biotechnology, Dallas, TX, USA; anti-Galectin-3—1:1000, #89572, Cell Signaling Technology, Denver, CO, USA; anti-ADAMTS5—1:500, ab41037, Abcam, Cambrige, UK; anti-GAPDH—1:1000, sc-25778, Santa Cruz Biotechnology, TX, USA.

Techniques: Western Blot, Transgenic Assay

Journal: Biomedical Journal

Article Title: Endothelial activation, cell-cell interactions, and inflammatory pathways in postoperative atrial fibrillation following cardiac surgery

doi: 10.1016/j.bj.2024.100821

Figure Lengend Snippet: Connexins expression and apoptosis level in a representative tissue sample from each type of patient: permanent AF, POAF and non-AF. Cx40: connexin 40; Cx43: connexin 43. Connexins can be seen in brown between the junctions of the cells. Apoptosis occurs when the nuclei of cardiomyocytes appear brown.

Article Snippet: After endogenous peroxidase and unspecific background blocking, sections were incubated with the corresponding primary antibody diluted in blocking solution overnight at 4 °C (anti-Cx40 polyclonal Antibody [36–4900, Invitrogen, dilution 1/100] and anti-Cx43 monoclonal Antibody [13–8300, Invitrogen, dilution 1/250]).

Techniques: Expressing

Journal: Biomedical Journal

Article Title: Endothelial activation, cell-cell interactions, and inflammatory pathways in postoperative atrial fibrillation following cardiac surgery

doi: 10.1016/j.bj.2024.100821

Figure Lengend Snippet: Connexin levels. A. Cx40. B. Cx43. ∗ p < 0.05 ∗∗ p < 0.01 ∗∗∗ p < 0.001.

Article Snippet: After endogenous peroxidase and unspecific background blocking, sections were incubated with the corresponding primary antibody diluted in blocking solution overnight at 4 °C (anti-Cx40 polyclonal Antibody [36–4900, Invitrogen, dilution 1/100] and anti-Cx43 monoclonal Antibody [13–8300, Invitrogen, dilution 1/250]).

Techniques: