Journal: Scientific Reports

Article Title: An exploratory in vitro co-culture of enteric neurons and smooth muscle cells demonstrates neuronal contribution to muscle layer formation

doi: 10.1038/s41598-026-39409-3

Figure Lengend Snippet: Schematic representation of plates with inserts and different layouts of muscle and ENS cells in a three-dimensional matrix: ( a ) plates with insert, which allows the medium to surround and support the co-culture from all sides, ( b ) SMCs without ENS cells in the 3D-matrix, ( c ) SMCs and ENS cells equally distributed in 3D-matrix, ( d ) SMCs distributed in upper and lower layers, ENS cells - in the middle layer of the 3D-matrix. ( e ) SMCs distributed in upper and lower layers, ENS cells densely arranged in the same plane of the middle layer of the 3D-matrix. ( f ) SMCs and ENS cells distributed in the middle layer of the 3D-matrix. ( g ) mixed SMCs and ENS cells densely arranged in the same plane of the middle layer of the 3D-matrix, ( h ) different layouts between SMCs and isolated myenteric plexus (ENS cells). SMCs alone, with direct contact and without direct contact to isolated myenteric plexus cells.

Article Snippet: Fig. 8 Schematic representation of plates with inserts and different layouts of muscle and ENS cells in a three-dimensional matrix: ( a ) plates with insert, which allows the medium to surround and support the co-culture from all sides, ( b ) SMCs without ENS cells in the 3D-matrix, ( c ) SMCs and ENS cells equally distributed in 3D-matrix, ( d ) SMCs distributed in upper and lower layers, ENS cells - in the middle layer of the 3D-matrix. ( e ) SMCs distributed in upper and lower layers, ENS cells densely arranged in the same plane of the middle layer of the 3D-matrix. ( f ) SMCs and ENS cells distributed in the middle layer of the 3D-matrix. ( g ) mixed SMCs and ENS cells densely arranged in the same plane of the middle layer of the 3D-matrix, ( h ) different layouts between SMCs and isolated myenteric plexus (ENS cells).

Techniques: Co-Culture Assay, Isolation

Journal: Scientific Reports

Article Title: An exploratory in vitro co-culture of enteric neurons and smooth muscle cells demonstrates neuronal contribution to muscle layer formation

doi: 10.1038/s41598-026-39409-3

Figure Lengend Snippet: Images of 3 weeks old co-culture of ENS cells and muscle cells in different layouts: ( a ) SMCs alone, ( b ) paracrine interaction between SMCs and ENS cells, ( c ) direct contact between SMCs and ENS cells Scale bars 100 μm, ( d ) paracrine interaction between SMCs and ENS in higher magnification, ( e ) Direct contact between smooth muscle and ENS cells in higher magnification.– muscle cells Scale bars 20 μm, ( f ) 3D reconstruction of muscle fibers in confocal microscopy by the direct contact between SMCs and ENS, ( j ) neurons, which are intercommunicated in the neuronal net within the muscle layer by the direct contact between SMCs and ENS. Confocal microscopy, Green (ß-Tubulin III) – neurons, red (smooth muscle actin (SMA)) – muscle cells. Scale bars 50 μm.

Article Snippet: Fig. 8 Schematic representation of plates with inserts and different layouts of muscle and ENS cells in a three-dimensional matrix: ( a ) plates with insert, which allows the medium to surround and support the co-culture from all sides, ( b ) SMCs without ENS cells in the 3D-matrix, ( c ) SMCs and ENS cells equally distributed in 3D-matrix, ( d ) SMCs distributed in upper and lower layers, ENS cells - in the middle layer of the 3D-matrix. ( e ) SMCs distributed in upper and lower layers, ENS cells densely arranged in the same plane of the middle layer of the 3D-matrix. ( f ) SMCs and ENS cells distributed in the middle layer of the 3D-matrix. ( g ) mixed SMCs and ENS cells densely arranged in the same plane of the middle layer of the 3D-matrix, ( h ) different layouts between SMCs and isolated myenteric plexus (ENS cells).

Techniques: Co-Culture Assay, Confocal Microscopy

Journal: Scientific Reports

Article Title: An exploratory in vitro co-culture of enteric neurons and smooth muscle cells demonstrates neuronal contribution to muscle layer formation

doi: 10.1038/s41598-026-39409-3

Figure Lengend Snippet: Imaging of the whole thickness of the 3D scaffolds after 3 weeks of ENS cells and muscle cells co-culture in different layouts: ( a ) SMCs without ENS cells, ( b ) paracrine interaction between SMCs and ENS cells, ( c ) direct contact between SMCs and ENS cells. Confocal microscopy, Green (ß-Tubulin III) – neurons, red (smooth muscle actin (SMA)) – muscle cells.

Article Snippet: Fig. 8 Schematic representation of plates with inserts and different layouts of muscle and ENS cells in a three-dimensional matrix: ( a ) plates with insert, which allows the medium to surround and support the co-culture from all sides, ( b ) SMCs without ENS cells in the 3D-matrix, ( c ) SMCs and ENS cells equally distributed in 3D-matrix, ( d ) SMCs distributed in upper and lower layers, ENS cells - in the middle layer of the 3D-matrix. ( e ) SMCs distributed in upper and lower layers, ENS cells densely arranged in the same plane of the middle layer of the 3D-matrix. ( f ) SMCs and ENS cells distributed in the middle layer of the 3D-matrix. ( g ) mixed SMCs and ENS cells densely arranged in the same plane of the middle layer of the 3D-matrix, ( h ) different layouts between SMCs and isolated myenteric plexus (ENS cells).

Techniques: Imaging, Co-Culture Assay, Confocal Microscopy

Journal: The Journal of Cell Biology

Article Title: Endothelial Msx1 transduces hemodynamic changes into an arteriogenic remodeling response

doi: 10.1083/jcb.201502003

Figure Lengend Snippet: LSS-dependent canonical BMP signaling triggers MSX1 expression in ECs. (A) HUAECs were transfected with control siRNA or siRNA against SMAD1/5 , seeded in fibronectin-coated chambers, and subsequently kept static or subjected for 1 h to an LSS of 20 dynes/cm 2 . MSX1 expression was analyzed and represented relative to the control static condition. n = 7. (B) Corresponding representative Western blot analysis for MSX1, pSMAD1/5/8, SMAD1, and GAPDH. (C) HUAECs were pretreated for 1 h with DMSO or 2.5-µM DMH1 and subjected to static or LSS (20 dynes/cm 2 ) conditions for 1 h in the presence of DMSO or DMH1. MSX1 expression was analyzed and represented relative to the DMSO condition. n = 3. (D–F) Cells transfected with control siRNA or siRNA against ALK6 ( n = 5), BMPR2 ( n = 6), or ENG ( n = 4) were exposed to static or flow conditions and were analyzed for MSX1 expression. *, P < 0.05 versus the indicated condition. (G) qRT-PCR analysis of HUAECs kept in static conditions or exposed to LSS (20 dynes/cm 2 for the indicated times). mRNA expression is represented relative to the static condition. n = 3. (A, C, and D–F) *, P < 0.05 versus static condition. ud, undetectable. (H) qRT-PCR analysis of HUAECs and SMCs serum starved for 1 h and exposed to 10 ng/µl BMP2, BMP6, or the corresponding amount of solute for 48 h. mRNA expression is represented relative to the control condition. n = 4 and 7. *, P < 0.05; #, P = 0.13 versus corresponding solute condition. Error bars represent the SEM.

Article Snippet: Freshly isolated HUAECs and uterine SMCs (Lonza) were seeded in 6-well plates at a density of 40,000 and 20,000 cells per well, respectively.

Techniques: Expressing, Transfection, Western Blot, Quantitative RT-PCR

Journal: The Journal of Cell Biology

Article Title: Endothelial Msx1 transduces hemodynamic changes into an arteriogenic remodeling response

doi: 10.1083/jcb.201502003

Figure Lengend Snippet: MSX1 induces an inflammatory proadhesive endothelial state. (A) Pictures of THP1 cells (in green) attached to HUAECs upon transduction with control Cherry or MSX1 plasmid for 3 d. Bars, 100 µm. (B) Quantification is represented as the number of monocytes per field of view. n = 6. *, P < 0.05 versus the Cherry control condition. (C) qRT-PCR expression analysis of ICAM1 and VCAM1 in MSX1 overexpression conditions mediated by plasmid transduction of HUAECs ( n = 10) or SMCs ( n = 3). *, P < 0.05 versus the Cherry control condition. (D) qRT-PCR analysis upon siRNA-mediated MSX1 knockdown. mRNA expression is represented relative to the control condition. n = 5–7. *, P < 0.05 versus the corresponding control condition. (E and F) qRT-PCR analysis on HUAECs transfected with control siRNA or siRNA against MSX1 seeded in fibronectin-coated chambers and subsequently kept under static conditions or subjected to 6 h of oscillatory flow. ICAM1 and VCAM1 expression is represented relative to the control static condition. n = 4 and n = 3, respectively. *, P < 0.05 versus the indicated condition. Error bars represent the SEM.

Article Snippet: Freshly isolated HUAECs and uterine SMCs (Lonza) were seeded in 6-well plates at a density of 40,000 and 20,000 cells per well, respectively.

Techniques: Transduction, Plasmid Preparation, Quantitative RT-PCR, Expressing, Over Expression, Transfection

Journal: Diabetologia

Article Title: Native incretins prevent the development of atherosclerotic lesions in apolipoprotein E knockout mice

doi: 10.1007/s00125-011-2241-2

Figure Lengend Snippet: Expression and abundance of GLP-1R and GIPR in monocytes/macrophages and arterial SMCs. a Glp1r and ( b ) Gipr mRNA in exudate peritoneal macrophages and tissues as labelled from non-treated Apoe −/− mice were measured by real-time RT-PCR. c Western blotting analyses of GLP-1R and ( d ) GIPR abundance in exudate peritoneal macrophages of non-treated Apoe −/− mice and other mouse tissues as labelled. Three independent experiments were performed. e Immunostaining of GLP-1R or GIPR in exudate peritoneal macrophages from non-treated Apoe −/− mice. Green, GLP-1R or GIPR as indicated; red, nuclei; red + green, overlay of GLP-1R or GIPR and nuclei. Scale bars 50 μm. f GLP1R and ( g ) GIPR mRNA levels in human coronary artery SMCs, THP1 cells and THP1-derived macrophages. h GIPR mRNA level in human monocytes and monocyte-derived macrophages were measured by real-time RT-PCR. Results were obtained from three to six independent experiments

Article Snippet: The expression of GIPR mRNA in human monocytes was far higher than in human macrophages and human coronary artery SMCs (Lonza) (Fig. ).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunostaining, Derivative Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: MicroRNA‐92a promotes vascular smooth muscle cell proliferation and migration through the ROCK/MLCK signalling pathway

doi: 10.1111/jcmm.14274

Figure Lengend Snippet: Expression of KLF4 in blood vessel walls of AS mice and effects of siRNA‐KLF4 (siKLF4) on vascular smooth muscle cells (VSMCs) function. A, Immunofluorescence staining for KLF4 (red) and α‐SMA (green) in AS mice and ML‐7 treatment mice. Nuclei were stained with DAPI (blue). Bars, 50 μm. B, The proliferation of siKLF4‐transfected Rat Primary Aortic SMCs were measured by CCK8 with 10 ng/mL PDGF‐BB stimulation. C and D, The migration of siKLF4‐transfected Rat Primary Aortic SMCs (C) and A7r5 (D) were measured by Boyden Chamber assay with 10 ng/mL PDGF‐BB stimulation. Migrated cells in each high‐power field (HPF, 400×) were quantitated and the results are shown on the right. E and F, The cells proliferation and migration of HASMCs were measured by CCK8 (E) and Boyden Chamber assay (F) respectively. Migrated cells in each high‐power field (HPF, 400×) were quantitated and the results are shown on the right. Data are presented as mean ± SD from three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Human aortic SMCs (HASMCs) were purchased from Chi Scientific Inc (Catalog No. 7‐1562, Jiangsu, China).

Techniques: Expressing, Immunofluorescence, Staining, Transfection, Migration, Boyden Chamber Assay