Journal: Investigative Ophthalmology & Visual Science

Article Title: MicroRNA 9 Is a Regulator of Endothelial to Mesenchymal Transition in Diabetic Retinopathy

doi: 10.1167/iovs.64.7.13

Figure Lengend Snippet: Primer Sequences Used for RT-qPCR

Article Snippet: Pecam1 (ab204527; Abcam, Cambridge, MA, USA) and Acta2 (NBP2-66429; Novus Biologicals, Littleton, CO, USA) ELISA kits were used according to the suppliers’ instructions.

Techniques: Sequencing

Journal: Investigative Ophthalmology & Visual Science

Article Title: MicroRNA 9 Is a Regulator of Endothelial to Mesenchymal Transition in Diabetic Retinopathy

doi: 10.1167/iovs.64.7.13

Figure Lengend Snippet: MiR-9 prevented diabetes-induced EndMT and diabetes-induced vascular leakage in mouse retinas. Streptozotocin-induced diabetes significantly reduced retinal mRNA expressions of endothelial markers ( A ) Pecam1 and ( B ) Cdh5 , and increased expressions of mesenchymal markers ( C ) Acta2 and ( D ) S100a4 in male WT mice. Such changes were prevented in diabetic M9 mice. Protein expressions of ( E ) Pecam1 and ( F ) Acta2 also followed the same pattern. ( G ) IgG staining was seen within the retinal capillaries of all mice ( arrow ). Diabetes caused leakage of IgG into the retinal layers of WT diabetic mice, resulting in intense (+++) staining throughout the retina when compared with the nondiabetic mice (+). Diabetic M9 mice showed minimal leakage of IgG into the retina and had staining intensity comparable to non-diabetic M9 mice (+). ND, nondiabetic; D, diabetic; M9, miR-9 transgenic (n = 6/group for mRNA, n = 3/group for protein, and n = 3/group for immunohistochemistry; molecular data normalized to WT-ND, mRNA data presented as ratio to β-actin mRNA, and protein data presented as ratio to total protein; * P < 0.05).

Article Snippet: Pecam1 (ab204527; Abcam, Cambridge, MA, USA) and Acta2 (NBP2-66429; Novus Biologicals, Littleton, CO, USA) ELISA kits were used according to the suppliers’ instructions.

Techniques: Staining, Transgenic Assay, Immunohistochemistry

Journal: Scientific reports

Article Title: Exploring the Endothelin-1 pathway in chronic thromboembolic pulmonary hypertension microvasculopathy.

doi: 10.1038/s41598-024-79623-5

Figure Lengend Snippet: Fig. 1. The Endothelin-1 (ET-1) Pathway Demonstrates Heightened Expression in Both the Plasma and Lung Microvessels of Chronic Thromboembolic Pulmonary Hypertension (CTEPH) Patients. (A) The concentration of ET-1 in venous plasma is elevated in CTEPH patients (n = 59) compared to control subjects (n = 41). (B) Co-immunofluorescent staining for ET-1 (red), von Willebrand factor (vWF, white), alpha- smooth muscle actin (α-SMA, green), and DAPI (blue) within lung microvessels from control subjects, CTEPH patients, and idiopathic pulmonary arterial hypertension (iPAH) patients, and the quantification of ET-1 fluorescent mean intensity (FMI) in the pulmonary endothelium (n = 5). (C) Co-immunofluorescent staining for ET-1 receptor A (ETA) (red), α-SMA (green), and DAPI (blue) in controls, CTEPH, and iPAH lungs, along with quantification of ETA FMI in pulmonary artery smooth muscle cells (PA-SMCs) (n = 5). (D) Co-immunofluorescent staining for phospho-myosin light chain (p-MLC) (red), α-SMA (green), and DAPI (blue) in controls, CTEPH, and iPAH lungs, along with quantification of p-MLC FMI in PA-SMCs (n = 5). All data are presented as mean with standard error of the mean (SEM). Statistical significance is denoted as * for P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001 and “ns” for not significant. The scale bar in all staining images is set at 100 µm.

Article Snippet: Immunofluorescent staining for ET-1 (Abcam for human lung staining, Abcam and Origen for piglet lung staining), ETA (Novus Biologicals for human staining, Abcam for piglet model staining), phosphorylated MLC (Cell signaling), alpha-smooth muscle actin (α-SMA, Novus Biologicals) were performed in human and piglet lung paraffin sections.

Techniques: Expressing, Clinical Proteomics, Concentration Assay, Control, Staining

Journal: Scientific reports

Article Title: Exploring the Endothelin-1 pathway in chronic thromboembolic pulmonary hypertension microvasculopathy.

doi: 10.1038/s41598-024-79623-5

Figure Lengend Snippet: Fig. 4. The Endothelin-1 (ET-1) Pathway Demonstrates Increased Expression in both the Plasma and Lung Microvessels of the CTEPH Piglet Model. (A) The concentration of ET-1 in venous plasma in the CTEPH piglet model compared to sham controls (n = 11). (B) Co-immunofluorescent staining for ET-1 (red), von Willebrand factor (vWF, white), alpha-smooth muscle actin (α-SMA, green), and DAPI (blue) within lung microvessels from the sham controls, non-obstructed (RSL), and obstructed (LSL) territories, along with quantification of fluorescent mean intensity (FMI) in the pulmonary endothelium (n = 5). (C) Co- immunofluorescent staining for ET-1 receptor A (ETA) (red), a-SMA (green), and DAPI (blue) in sham lungs, and the RSL and LSL territories, along with quantification of FMI in pulmonary artery smooth muscle cells (PA-SMCs) (n = 5). (D) Co-immunofluorescent staining for phospho-myosin light chain (p-MLC) (red), α-SMA (green), and DAPI (blue) in sham lungs, and the RSL and LSL territories, along with quantification of FMI in PA- SMCs (n = 5). All data are presented as mean values with standard error of the mean (SEM). Statistical significance is denoted as * for P < 0.05, **P < 0.01, and ***P < 0.001. The scale bar in all staining images is set at 100 µm. LSL: left superior lobe; RSL: right superior lobe.

Article Snippet: Immunofluorescent staining for ET-1 (Abcam for human lung staining, Abcam and Origen for piglet lung staining), ETA (Novus Biologicals for human staining, Abcam for piglet model staining), phosphorylated MLC (Cell signaling), alpha-smooth muscle actin (α-SMA, Novus Biologicals) were performed in human and piglet lung paraffin sections.

Techniques: Expressing, Clinical Proteomics, Concentration Assay, Staining